Camptothecin analogues, preparation methods therefor, use thereof as drugs, and pharmaceutical compositions containing said analogues

ABSTRACT

A camptothecin analog characterized in that the hydroxy lactone of the camptothecin is a β-hydroxy lactone or the corresponding β-hydroxyacid, resulting from the opening of said lactone, or a derivative of said β-hydroxyacid, or a Pharmaceutically acceptable salt thereof, is disclosed. In particular, compounds of formulae (I) and (II) are disclosed. Methods for preparing the compounds of formulae (I) and (II), pharmaceutical compositions containing said containing said compounds, and their use, particularly as topoisomerase inhibitors and antitumoral drugs, are also disclosed.

Camptothecin is a natural compound that was isolated for the first timefrom the leaves and the bark of the Chinese plant called camptothecaacuminata (see Wall et al., J. Amer. Chem. Soc. 88: 3888 (1966)).Camptothecin is a pentacyclic compound comprised of an indolizino[1,2-b] quinoline fragment fused with a six chained α-hydroxylactone.The carbon in position 20, which carries the α-hydroxy, is asymmetricand confers a rotational capacity to the molecule. The natural form ofcamptothecin possesses the absolute "S" configuration to the carbon 20and complies with the following formula: ##STR2## Camptothecin presentsan anti-proliferating activity in several cancerous cellular lines,comprising the cellular lines of human tumors of the colon, of the lungand of the breast (Suffness, M. et al., The Alkaloids Chemistry andPharmacology, Bross, A., ed., Vol. 25, p. 73 (Academic Press, 1985)). Itis suggested that the anti-proliferating activity of camptothecin is inrelation to its inhibiting activity of topoisomerase I of DNA.

It was indicated that the α-hydroxylactone was an absolute necessityboth for in vivo and in vitro activity of the camptothecin(Camptothecins: New Anticancer Agents, Putmesil, M. et al., ed., p. 27(CRC Press. 1995); Wall, M. et al., Cancer Res. 55: 753 (1995);Hertzberg et al., J. Med. Chem. 32: 715 (1982) and Crow et al., J. Med.Chem. 35: 160 (1992)). The present invention concerns a new class ofcamptothecin compounds, in which a β-hydroxylactone replaces the naturalα-hydroxylactone of the camptothecin. The compounds according to thepresent invention exhibit powerful biological activity which isunexpected with respect to prior art.

Thus the invention has as its object new compounds of camptothecin whichdiffer from all known compounds of camptothecin, in the sense that theycontain a β-hydroxylactone (or its open hydroxycarboxylic form) in placeof an α-hydroxylactone (or its open hydroxycarboxylic form); or apharmaceutically acceptable salt of the latter. By compound ofcamptothecin it is understood a compound presenting the same structuralskeleton as that of camptothecin (that is to say an indolizino [1,2-b]quinoline fragment that is fused to a six chained α-hydroxylactone),with or without other chemical substitutions on the skeletal structure(for example the camptothecin compounds). Different camptothecincompounds are well known by specialists, as described below. Byβ-hydroxylactone is meant a lactone that is comprised of a supplementarycarbon atom between the carbon of the carboxyl and the α-carbon carryingthe hydroxy group on the α-hydroxylactone. The β-hydroxylactone of seven"closed" or "open" chains, in which the ester bond between the carbonylgroup and the adjacent oxygen atom is hydrolized, which shows by theformation of a carboxylic acid group and a hydroxyl group, which groupsmay or may not be substituted.

A compound of camptothecin according to the invention may then comprisesubstitutions on the indolizino [1,2-5] quinoline fragment (for exampleto improve the solubility of the compound) or on the open or closedβ-hydroxylactone (for example to improve the stability of the compound).The examples of substitutions on closed β-hydroxylactone are comprisedof an alkyl substitution (for example ethyl) on the β-carbon. Theexamples of substitution on the open β-hydroxylactone are comprised ofan alkyl substitution on the β-carbon, substitutions (for example anamidation) on the carboxylic acid result, and the substitutions (forexample an esterification) or suppressions of a hydroxyl group result.

The invention has more particularly for subject matter compounds offormula (I) and of formula (II), ##STR3## in racemic form, enantiomer orany combination of these forms, in which R₁ represents a lower alkyl, alower alkenyl, a lower alkynyl, a lower haloalkyl, and lower alkoxylower alkyl or a lower alkylthio lower alkyl;

R₂, R₃, and R₄ represent, independently, H, halo, lower halo alkyl,lower alkyl, lower alkenyl, cyano, lower cyano alkyl, nitro, lower nitroalkyl, amido, lower amido alkyl, hydrazino, lower hydrazino alkyl,azido, lower azido alkyl, (CH₂)_(m) NR₆ R₇, (CH₂)_(m) OR₆, (CH₂)_(m)SR₆, (CH₂)_(m) CO₂ R₆, (CH₂)_(m) NR₆ C(O)R₈,(CH₂)_(m) C(O)R₈, (CH₂)_(m)OC(O)R₈, O(CH₂)_(m) NR₆ R₇, OC(O)NR₆ R₇, OC(O)(CH₂)_(m) CO₂ R₆, or(CH₂)_(n) [N═X], OC(O)[N═X], (CH₂)_(m) OC(O)[N═X] (in which [N═X], inthis invention represents a heterocyclic group of 4 to 7 chains with thenitrogen atom N, which is a member of the heterocyclic group, and Xrepresents the resulting members, necessary to complete the heterocyclicgroup, selected from among the group comprised of O, S, CH₂, CH, N, NR₉and COR₁₀), aryl or lower aryl alkyl substituted (that is to say,substituted one to four times on the heterocyclic or aryl group) or notsubstituted, in which the substituent is a lower alkyl, halo, nitro,amino, lower alkylamino, lower haloalkyl, lower hydroxy alkyl, loweralkoxy, or lower alkoxy lower alkyl) or R₂ and R₃ form together a chainof 3 or 4 links, in which the elements of the chain are selected fromamong the group comprised of CH, CH₂, O, S, N or NR₉ ;

R₅ represents H, halo, lower halo alkyl, lower alkyl, lower alkoxy,lower alkoxy lower alkyl, lower alkylthio lower alkyl, cycloalkyl, lowercycloalkyl alkyl, cyano, cyano alkyl, lower alkyl lower sulphonyl alkyl,lower hydroxy alkyl, nitro, (CH₂)_(m) C(O)R₈, (CH₂)_(m) NR₆ C(O)R₈,(CH₂)_(m) NR₆ R₇, (CH₂)_(m) N(CH₃)(CH₂)_(n) NR₆ R₇, CH₂)_(m) OC(O)R₈,(CH₂)_(m) OC(O)NR₆ R₇, (CH₂)_(m) S(O)_(q) R₁₁, (CH₂)_(m) P(O)R₁₂ R₁₃,(CH₂)₂ P(S)R₁₂ R₁₃, or (CH₂)_(n) [N═X], OC(O)[N═X], (CH₂)_(m)OC(O)[N═X], aryl or substituted lower aryl alkyl (that is to say one tofour times on the aryl group or heteroaryl) or not substituted, in whichthe substituent is a lower alkyl, halo, nitro, amino, lower alkyl amino,lower halo alkyl, lower hydroxy alkyl, lower alkoxy or lower alkoxylower alkyl;

R₆ and R₇ represent, independently, H, and lower alkyl, lower hydroxyalkyl, lower alkyl lower amino alkyl, lower amino alkyl, cycloalkyl,lower cycloalkyl alkyl, lower alkenyl, lower alkoxy lower alkyl, lowerhalo alkyl, or aryl or substituted lower aryl alkyl (that is to say oneto four times on the aryl group) or non-substituted, in which thesubstituent is a lower alkyl, halo, nitro, amino, lower alkyl amino,lower halo alkyl, lower hydroxy alkyl, lower alkoxy, or lower alkoxylower alkyl;

R₈ represents H, and lower alkyl, lower hydroxy alkyl, amino, loweralkyl amino, lower alkyl lower amino alkyl, lower amino alkyl,cycloalkyl, lower cycloalkyl alkyl, lower alkenyl, lower alkoxy, loweralkoxy lower alkyl, lower halo alkyl, or aryl or substituted lower arylalkyl (that is to say, one to four times on the aryl group) ornon-substituted, in which the substituent is a lower alkyl, halo, nitro,amino, lower alkyl amino, lower halo alkyl, lower hydroxy alkyl, loweralkoxy, or lower alkoxy lower alkyl;

R₉ represents H, a lower alkyl, lower halo alkyl, aryl, or substitutedaryl by one or more groups chosen from among the lower radical alkyl,halo, nitro amino, lower alkyl amino, lower halo alkyl, lower hydroxyalkyl, lower alkoxy, or lower alkoxy lower alkyl;

R₁₀ represents H, a lower alkyl, lower halo alkyl, lower alkoxy, aryl orsubstituted aryl (that is to say, presenting one to four substituents onthe aryl group) by one or more groups chosen from among the lowerradical alkyl, lower halo alkyl, lower hydroxy alkyl, or lower alkoxylower alkyl;

R₁₁ represents a lower alkyl, aryl, (CH₂)_(m) OR₁₄, (CH₂)_(m) SR₁₄,(CH₂)₂ NR₁₄ R₁₅ or (CH₂)_(m) [N═X];

R₁₂ and R₁₃ represent, independently, a lower alkyl, aryl, lower alkoxy,aryloxy or amino;

R₁₄ and R₁₅ represent, independently, H, and lower alkyl or aryl;

R₁₆ represents H or OR₂₁ ;

R₁₇ represents OR₆ or NR₆ R₇ ;

R₁₈ and R₁₉ represent, independently, H, halo, lower alkyl, lower alkoxyor hydroxy;

R₂₀ represents H or halo;

R₂₁ represents H, a lower alkyl, CHO or C(O)(CH₂)_(m) CH₃ ;

m is an integer comprised between 0 and 6;

n is 1 or 2; and

q represents an integer from 0 to 2; and [N═X] represents a heterocyclicgroup with 4 to 7 links, X representing the necessary chain to completesaid heterocyclic group and selected from the group comprised of O, S,CH₂, CH, N, NR₉ and COR₉ ; or a pharmaceutically acceptable salt of thelatter.

The invention has particularly as its subject matter the compounds offormula I and II such as defined above, in which R₁ represents a loweralkyl, lower alkenyl, lower halo alkyl, lower alkoxy lower alkyl orlower alkylthio lower alkyl; R₅ represents H, a halo, lower halo alkyl,lower alkyl, lower alkoxy lower alkoxy lower alkyl, lower alkylthiolower alkyl, cycloalkyl, lower cycloalkyl alkyl, cyano, cyano alkyl,lower hydroxy alkyl, nitro, (CH₂)_(m) C(O)R₈, (CH₂)_(m) NR₆ C(O)R₈,(CH₂)_(m) NR₆ R₇, (CH₂)_(m) N(CH₃)(CH₂)_(n) NR₆ R₇, (CH₂)_(m) OC(O)R₈,(CH2)_(m) OC(O)NR₆ R₇, or (CH₂)_(n) [N═X], OC(O)[N═X], (CH₂)_(m)OC[N═X], aryl or lower aryl alkyl, substituted or not substituted; R₁₂and R₁₃ represent, independently, a lower alkyl; R₁₆ represents OR₂₁ ;and R₁₈, R₁₉ and R₂₀ represent H.

The invention has more particularly as its subject matter the compoundsof formula I and II as is defined above in which R₁ represents a loweralkyl, lower alkenyl, lower halo alkyl or lower alkoxy lower alkyl; R₂,R₃ and R₄ represent, independently, H, halo, lower halo alkyl, loweralkyl, nitro, amido, lower amido alkyl, hydrazino, lower hydrazinoalkyl, azido, lower azido alkyl, (CH₂)_(m) NR₆ R₇, (CH₂)_(m) OR₆,(CH₂)_(m) SR₆, (CH₂)_(m) C(O)R₈, OC(O)NR₆ R₇, (CH₂)_(n) [N═X], or(CH₂)_(m) OC(O)[N═X] substituted or non-substituted, or OC(O)[N═X]; orR₂ and R₃, together form a chain of 3 or 4 links, in which said elementsof the chain are chosen from the group comprised of CH, CH₂, O, S, N, orNR₉ ; R₅ represents H, halo, lower halo alkyl, lower alkyl, loweralkoxy, lower alkoxy lower alkyl, lower alkylthio lower alkyl, lowerhydroxy alkyl, nitro, (CH₂)_(m) C(O)R₈, (CH₂)_(m) NR₆ C(O)R₈, (CH₂)_(m)NR₆ R₇, (CH₂)_(m) N(CH₃)(CH₂)_(n) NR₆ R₇, (CH₂)_(m) OC(O)R₈, (CH₂)_(m)OC(O)NR₆ R₇, (CH₂)_(n) [N═X] or OC(O)[N═X] substituted ornon-substituted, or (CH₂)_(m) OC(O)[N═X]; R₆ and R₇ represent,independently, H, a lower alkyl, lower hydroxy alkyl, lower alkyl loweramino alkyl, lower amino alkyl, cycloalkyl, lower cycloalkyl alkyl,lower alkoxy lower alkyl, aryl, lower aryl alkyl or lower halo alkyl; R₈represents H, and lower alkyl, lower hydroxy alkyl, lower alkyl amino,lower alkyl lower amino alkyl, lower amino alkyl, cycloalkyl, lowercycloalkyl alkyl, lower alkenyl, lower alkoxy, lower alkoxy lower alkyl,lower halo alkyl, aryl or lower aryl alkyl; R₉ represents H, a loweralkyl, or a lower halo alkyl; R₁₀ represents H, and lower alkyl, lowerhalo alkyl, or a lower alkoxy; R₁₁ represents and lower alkyl; and R₁₄and R₁₅ represent, independently, H or a lower alkyl.

The invention has more particularly as its subject matter the compoundsof formula I such as is defined in which R₁ represents the ethyl group;R₂ and R₃ represent, independently, H, a lower alkyl, halo, lower haloalkyl or (CH₂)_(m) OR₆, or R₂ and R₃, together, form a methylenedioxy oran ethylenedioxy; and R₄ and R₅ represent, independently, H, and loweralkyl, (CH₂)_(m) NR₆ R₇ or (CH₂)_(n) [N═X] non-substituted orsubstituted by a lower alkyl. Preferably, R₄ represents H or (CH₂)_(m)NR₆ R₇ in which R₆ and R₇ represent, independently, H or a lower alkyl,and R₅ represents H, and lower alkyl or (CH₂)_(n) [N═X] substituted ornon-substituted by a lower alkyl; as example of [N═X] substituted ornon-substituted, the piperidyl radical, morpholinyl, piperazinyl,imidazolyl and 4-methylpiperazinyl may be cited.

In a manner more preferable, R₂ represents H or a halo, and preferablyH, chloro or fluoro; and R₃ represents H, a lower alkyl, halo or OR₆ inwhich R₆ represents H, a lower alkyl or a lower aryl alkyl, andpreferably H, fluoro, chloro, methyl or methoxy. In an equallypreferable manner, R₂ and R₃ form together a methylenedioxy or anethylenedioxy.

More particularly, the invention has as its subject matter the productsdescribed hereafter in the examples, in particular the productscorresponding to the following formulas:

5-ethyl-4,5-dihydro-5-hydroxy-1H-oxepino [3',4':6,7]-indolizino [1,2-b]quinoline-3,15 (4H, 13H)-dione

5,12-diethyl-4,5-dihydro-5-hydroxy-1H-oxepino [3',4': 6,7]-indolizino[1,2-b] quinoline-3,15 (4H,13H)-dione

8-ethyl-2,3,8,9-tetrahydro-8-hydroxy-10H,12H-[1,4]dioxino [2,3-g]oxepino [3',4':6,7] indolizino [1,2-b] quinoline-10,13 (15H)-dione

10-benzyloxy-5-ethyl-4,5-dihydro-5-hydroxy-1H-oxepino[3',4':6,7]-indolizino [1,2-b] quinoline-3,15 (4H,13H)-dione

5-ethyl-4,5-dihydro-5,10,-dihydroxy-1H-oxepino [3',4':6,7]-indolizino[1,2-b] quinoline-3,15 (4H,13H)-dione

11-(dimethylamino)methyl-5-ethyl-4,5-dihydro-5,10-dihydroxy-1H-oxepino[3',4':6,7]-indolizino [1,2-b] quinoline-3,15 (4H,13H)-dione

5-ethyl-9-fluoro-4,5-dihydro-5-hydroxy-10-methoxy-1H-oxepino [3',4':6,7]indolizino [1,2-b] quinoline-3,15(4H,13H)-dione

9-chloro-5-ethyl-4,5-dihydro-5-hydroxy-10-methyl-1H-oxepino [3',4':6,7]indolizino [1,2-b] quinoline-3,15(4H,13H)-dione

5-ethyl-9,10-difluoro-4,5-dihydro-5-hydroxy-1H-oxepino [3',4':6,7]indolizino [1,2-b] quinoline-3,15(4H,13H)-dione

7-ethyl-7,8-dihydro-7-hydroxy-9H,11H-[1,3] dioxole [4,5-g] oxepino [3',4':6,7] indolizino [1,2-b] quinoline-9,12(14H)-dione

9-chloro-5-ethyl-4,5-dihydro-5-hydroxy-10-methoxy-1H-oxepino [3',4':6,7]indolizino [1,2-b] quinoline-3,15(4H,13H)-dione

5-ethyl-4,5-dihydro-5-hydroxy-10-methoxy-1H-oxepino [3',4':6,7]indolizino [1,2-b] quinoline-3,15(4H,13H)-dione

9,11-dichloro-5-ethyl-4,5-dihydro-5-hydroxy-1H-oxepino [3',4':6,7]indolizino [1,2-b] quinoline-3,15(4H,13H)-dione

5-ethyl-9-fluoro-4,5-dihydro-5-hydroxy-10-methyl-1H-oxepino [3',4':6,7]indolizino [1,2-b] quinoline-3,15 (4H,13H)-dione

5-ethyl-10-fluoro-4,5-dihydro-5-hydroxy-1H-oxepino [3',4':6,7]indolizino [1,2-b] quinoline-3,15(4H,13H)-dione

10-chloro-5-ethyl-4,5-dihydro-5-hydroxy-1H-oxepino[3',4':6,7] indolizino[1,2-b] quinoline-3,15(4H,13H)-dione

10-chloro-5-ethyl-9-fluoro-4,5-dihydro-5-hydroxy-1H-oxepino [3',4':6,7]indolizino [1,2-b] quinoline-3,15(4H,13H)-dione

5,12-diethyl-4,5-dihydro-5,10-dihydroxy-11-morpholino methyl-1H-oxepino[3',4':6,7] indolizino [1,2-b] quinoline-3,15(4H,13H)-dione

5,12-diethyl-9-fluoro-4,5-dihydro-5-hydroxy-10-methoxy-1H-oxepino[3',4':6,7] indolizino [1,2-b] quinoline-3,15(4H,13H)-dione

5-ethyl-4,5-dihydro-5-hydroxy-12-methyl-1H-oxepino [3',4':6,7]indolizino[1,2-b] quinoline-3,15 (4H,13H)-dione

9-chloro-5-ethyl-4,5-dihydro-5-hydroxy-10-methoxy-12-(4-methylpiperazinomethyl)-1H-oxepino [3',4':6,7] indolizino [1,2-b]quinoline-3,15(4H,13H)-dione

9-chloro-5-ethyl-4,5-dihydro-5-hydroxy-10-methoxy-12-morpholinomethyl-1H-oxepino[3',4':6,7] indolizino [1,2-b] quinoline-3,15(4H,13H)-dione

5-ethyl-4,5-dihydro-5-hydroxy-12-(4-methyl piperazinomethyl)-1H-oxepino[3',4':6,7] indolizino [1,2-b] quinoline-3,15(4H,13H)-dione

5-ethyl-4,5-dihydro-5-hydroxy-12-piperidinomethyl-1H-oxepino [3',4':6,7]indolizino [1,2-b] quinoline-3,15(4H,13H)-dione

5-ethyl-4,5-dihydro-5-hydroxy-12-morpholinomethyl-1H-oxepino [3',4':6,7] indolizino [1,2-b] quinoline-3,15(4H,13H)-dione

5-ethyl-10-fluoro-4,5-dihydro-5-hydroxy-12-(4-methylpiperazinomethyl)-1H-oxepino[3',4':6,7] indolizino [1,2-b] quinoline-3,15(4H,13H)-dione

5-ethyl-10-fluoro-4,5-dihydro-5-hydroxy-12-morpholinomethyl-1H-oxepino[3',4':6,7] indolizino [1,2-b] quinoline-3,15(4H,13H)-dione

5-ethyl-9-fluoro-4,5-dihydro-5-hydroxy-10-methyl-12-(4-methylpiperazinomethyl)-1H-oxepino[3',4':6,7] indolizino [1,2-b] quinoline-3,15(4H,13H)-dione

5-ethyl-9-fluoro-4,5-dihydro-5-hydroxy-10-methyl-12-morpholinomethyl-1H-oxepino[3',4':6,7] indolizino [1,2-b] quinoline-3,15(4H,13H)-dione

5-ethyl-9-fluoro-4,5-dihydro-5-hydroxy-10-methyl-12-piperidinomethyl-1H-oxepino[3',4':6,7] indolizino [1,2-b] quinoline-3,15(4H,13H)-dione

8-ethyl-2,3,8,9-tetrahydro-8-hydroxy-16-(4-methyl piperazinomethyl)-10H,12H-(1,4) dioxino (2,3-g) oxepino [3',4':6,7] indolizino [1,2-b]quinoline-10,13[15H]-dione

9-chloro-5-ethyl-10-fluoro-4,5-dihydro-5-hydroxy-12-morpholinomethyl-1H-oxepino[3',4':6,7] indolizino [1,2-b] quinoline-3,15(4H,13H)-dione

or a pharmaceutically acceptable salt of the latter.

The invention has more particularly for object equally the compounds offormula II as is described above, in which R₁ represents the ethylgroup; R₂ and R₃ represent, independently, H, and lower alkyl, halo,lower halo alkyl, or (CH₂)_(m) OR₆, or R₂ and R₃ form together amethylenedioxy, or an etnylenedioxy; R₄ and R₅ represent, independently,H, and lower alkyl, (CH₂)_(m) NR₆ R₇, or (CH₂)_(n) [N═X] non-substitutedor substituted by a lower alkyl; R₂₀ represents H and R₁₇ representsOR₆, in which R₆ represents H or a lower alkyl, or NR₆ R₇ in which R₆and R₇, independently represent H, and lower alkyl, aryl or lower arylalkyl. Preferably R₄ represents H or (CH₂)_(m) NR₆ R₇, in which R₆ andR₇ represent, independently, H or a lower alkyl; R₅ represents H, alower alkyl, or (CH₂)_(n) [N═X] non-substituted or substituted by alower alkyl and R₁₇ represents OR₆ in which R₆ represents H or a loweralkyl; or a pharmaceutically acceptable salt of the latter. As exampleof [N═X] the piperidyl radical, morpholinyl, piperazinyl, imidazolyl and4-methylpiperazinyl may be cited.

Even more preferable, R₂ represents H or halo and preferably H, chloroor fluoro; R₃ represents H, a lower alkyl, halo or OR₆ in which R₆represents H, a lower alkyl or a lower aryl alkyl and preferably H,fluoro, chloro, methyl or methoxy. Equally as preferential, R₂ and R₃form together methylenedioxy or ethylenedioxy.

More particularly, the invention has for object the products describedhereafter in the examples, in particular the products responding to thefollowing formulas:

tert-butyl β-ethyl-β-hydroxy-γ-(8-hydroxymethyl-9-oxo(11H)-indolizino-[1,2-b] quinoline-7-yl)-propionate

ethyl β-ethyl-β-hydroxy-γ-(8-hydroxymethyl-9-oxo(11H)-indolizino-[1,2-b] quinoline-7-yl)-propionate

β-ethyl-β-hydroxy-γ-(8-hydroxymethyl-9-oxo (11H)-indolizino-[1,2-b]quinoline-7-yl)-propionic acid

methyl β-ethyl-β-hydroxy-γ-(8-hydroxymethyl-9-oxo(11H)-indolizino-[1,2-b] quinoline-7-yl)-propionate

ethyl β-ethyl-α,α-difluoro-β-hydroxy-γ-(8-hydroxymethyl-9-oxo(11H)-indolizino-[1,2-b] quinoline-7-yl)-propionate

ethyl β-ethyl-β-hydroxy-γ-(8-hydroxymethyl-9-oxo(11H)-indolizino-[1,2-b] quinoline-7-yl)-propionate

tert-butyl β-ethyl-β-hydroxy-γ-(8-hydroxymethyl-9-oxo(11H)-indolizino-[1,2-b] quinoline-7-yl)-propionate

β-ethyl-γ-(12-ethyl-8-hydroxymethyl-9-oxo (11H)-indolizino-[1,2-b]quinoline-7-yl)-β-hydroxy-propionic acid

γ-(12-benzyloxy-8-hydroxymethyl-9-oxo (11H)-indolizino [1,2-b]quinoline-7-yl)-β-ethyl-β-hydroxy-propionic acid (E)

or a pharmaceutically acceptable salt of the latter.

Such as it is used here, the term lower in reference to alkyl, alkylthioand alkoxy groups designate the saturated aliphatic hydrocarbon groups,linear or branched, comprising 1 to 6 carbons, as for example methyl,ethyl, propyl, isopropyl, butyl, t-butyl, methylthio, ethylthio,methoxy, and ethoxy. In reference to the alkenyl or alkynyl groups, theterm lower designates the groups comprising 2 to 6 carbon atoms and oneor more double or triple bonds, as for example the vinyl, allyl,isopropenyl, pentenyl, hexanyl, propenyl ethynyl, propynyl and butynylgroups. The term cycloalkyl designates a cycle comprising 3 to 7 carbonatoms, as for example the cyclopropyl, cyclobutyl, cyclopentyl orcyclohexyl groups. The term aryl designates a hydrocarbon compoundmono-, di- or tricyclic with at least one aromatic cycle, each cyclecontaining a maximum of 7 links, as for example phenyl, naphthyl,anthracyl, biphenyl or indenyl. The term halo signifies chloro, bromo,iodo, or fluoro. The radicals corresponding to the expressions lowerhalo alkyl, lower cyano alkyl, lower nitro alkyl, lower amido alkyl,lower hydrazino alkyl, lower azido alkyl, lower aryl alkyl, lowerhydroxy alkyl, lower alkoxy lower alkyl, lower alkylthio lower alkyl,and lower alkyl sulphonyl lower alkyl are substituted, respectively, byone to three halo, cyano, nitro, amido, hydrazino, azido, aryl, hydroxy,lower alkoxy, lower alkylthio or lower sulphonyl groups. The lower alkylamino radical may contain one or two lower alkyl groups, and representfor example NHCH₃, NHCH₂ CH₃, N(CH₃)₂, or N(CH₃)(CH₂ CH₃). The examplesof [N═X] comprise the piperidinyl, morpholinyl, piperizinyl andimidazolyl groups.

As is observed for camptothecin, the carbon atom carrying the hydroxyfunction on the β-hydroxylactone or the α-hydroxycarboxylate group ofthe compounds, according to the present invention, is asymmetrical.Consequently, the compounds according to the present invention have twopossible enantiomer configurations, that is to say in the "R" and "S"configurations. The present invention includes the two enantiomerconfigurations and all the combinations of these configurations,comprising as well the "RS" racemic mixtures. In the interest ofsimplicity, when any specific configuration is not indicated in thestructural formulas, it is understood that the two enantiomerconfigurations and their mixtures are represented.

The invention has as well as its subject matter the preparationprocedures of the compounds of general formula I and II, either fromcamptothecin or from substituted camptothecin, or by total chemicalsynthesis.

The invention concerns thus a preparational procedure of formula I andII compounds, according to the invention starting from camptothecin orfrom substituted camptothecin characterized in that

α-hydroxylactone of the camptothecin is reduced to the general formula##STR4## in which R₁, R₂, R₃, R₄, R₅ and R₂₀ have the significanceindicated above, to obtain the α-hydroxylactol of general formula A##STR5## in which R₁, R₂, R₃, R₄, R₅ and R₂₀ have the significanceindicated above.

In compound A thus formed, the carbon-carbon bond bonding theneighboring carbinol are cut by treatment with an appropriate oxydizingagent so as to result in a formula B compound ##STR6## in which R₁, R₂,R₃, R₄, R₅ and R₂₀ have the significance indicated above.

It is subsequently treated with a functionalized alkylizing agent andthe formyl functional group of the formula B compound is cut to give aβ-hydroxyester of general formula C ##STR7## in which R₁, R₂, R₃, R₄,R₅, R₁₈, R₁₉, and R₂₀ have the significance indicated above and R₁₇represents OR₆ and R₆ represents a lower alkyl, cycloalkyl, lowercycloalkyl alkyl, lower alkenyl, lower alkoxy lower alkyl or aryl orlower aryl alkyl;

said compound of general formula C is cyclized resulting in theβ-hydroxylactonic compound of general formula D ##STR8## in which R₁,R₂, R₃, R₄, R₅, R₁₈, R₁₉ and R₂₀ have the significance indicated above,

the lactone of general formula D is opened, resulting in the formula Ecompound ##STR9## in which R₁, R₂, R₃, R₄, R₅, R₁₈, R₁₉ and R₂₀ have thesignificance indicated above; R₁₆ represents OR₂₁ in which R₂₁represents H or a lower alkyl; and R₁₇ represents OR₆ or NHR₆ and R₆represents H, a lower alkyl, cycloalkyl, lower cycloalkyl alkyl, loweralkenyl, lower alkoxy lower alkyl, or aryl or lower aryl alkyl.

Certain formula E compounds may be obtained as well by hydrolyzing theester function of the corresponding formula C compounds. The generalformula E compounds in which R₁₆ and/or R₁₇ represent, independently,the hydroxy radical, may be esterified or amidified under the classicconditions known by persons skilled in the art to obtain the esters orthe amides corresponding to formula E.

In the above process, the R₁, R₂, R₃ and R₄ groups may be protected ifnecessary according to the classical methods of protection (Greene, T.,Protective Groups in Organic Synthesis 10-86 (John Wiley & Sons 1981)).At the time of this procedure, the reduction occurs with the help of areducing agent in an appropriate solvent such as, for example, sodiumborohydride in methanol. The stage corresponding to the formation ofcompound B from compound A, is implemented under oxidizing conditionssuch as, for example, with lead tetraacetate, periodic acid or sodiummetaperiodate in an appropriate solvent such as, for example, aceticacid. The treatment by a functionalized alkylizing agent may beimplemented with the help of a metallic derivative for example lithiumor zinc, or with the help of a carboxylic ester in an anhydrous aproticsolvent such as for example tetrahydrofurane. The stage of lactonisationthat compound D to be obtained from compound C is generally carried outunder acidic conditions, for example, by treatment with trifluoroaceticacid or hydrochloric gas dissolved in an anhydrous solvent such asdichloromethane or dioxane. The opening of the lactonic cycle ofcompound D to obtain compound E, may be done, for example, byhydrolysing under alkaline conditions followed by neutralization.

The examples of substituted camptothecins, used as starting products,may be found in the U.S. Pat. Nos. 4,473,692, 4,604,463, 4,894,956,5,162,532, 5,395,939, 5,315,007, 5,264,579, 5,258,516, 5,254,690,5,212,317, and 5,341,745, the PCT Patent Application Nos. US91/08028,US94/06451, US90/05172, US92/04611, US93/10987, US91/09598, EP94/03058and EP95/00393 and the European Patent Applications Nos. 325 247, 495432, 321 122 and 540 099.

The invention concerns as well a preparation procedure of the compoundsof formula I and II, characterized in that

compound of general formula M is coupled ##STR10## in which R₁, R₁₈ andR₁₉ have the significance indicated above and R₂₀ represents hydrogen ora halogen atom, with a 2-halo-3-quinoline-methanol of general formula N##STR11## in which R₂, R₃, R₄ and R₅ have the significance indicatedabove and X represents a halogen atom, resulting in the formula Ocompound ##STR12## in which R₁, R₂, R₃, R₄, R₅, R₁₈, R₁₉, R₂₀ and X havethe significance indicated above:

then the general formula O compound is cyclized to obtain the formula Dcompound such as is defined above.

In the above method, the R₁, R₂, R₃ and R₄ groups may be protected ifnecessary according to the classical methods of protection (Greene, T.,Protective Groups in Organic Synthesis 10-86 (John Wiley & Sons 1981)).The formation of compound O from the general formula M and N compoundsis carried out by a treatment known by persons skilled in the art by thename of Mitsunobu reaction (refer to Mitsunobu, O. et al., Syntheses,p.1 (1981)). It is comprised of displacing the hydroxyl function ofcompound N by a nucleophile such as compound M, or a deprotonatedderivative of the latter, by treatment with a phosphine, for exampletriphenylphosphine, and azodicarboxylate derivative, for example diethylazodicarboxylate, in an aprotic solvent such as, for example,tetrahydrofuran or N,N-dimethylformamide. The cyclisation of compound Ooccurs preferably in the presence of a pallidin catalyst (for examplepalladium diacetate) under basic conditions (provided for example by analkaline acetate potentially combined with a phase transfer agent suchas for example tetrabutylammonium bromide), in an aprotic solvent suchas acetonitril or N,N-dimethylformamide, at a temperature between 50° C.and 120° C. (R. Grigg et al., Tetrahedron 46, page 4003 (1990)).

The general formula M compounds are new. They may be prepared accordingto a method characterized in that

the carbonyl is protected of a pyridine of general formula ##STR13## inwhich R₁ and R₂₀ have the significance indicated above and R₂₂represents a halogen atom or a lower alkoxy, by an acetal function,resulting in the general formula F compound ##STR14## in which R₁, R₂₀and R₂₂ have the significance indicated above and the Z and Z' groupsrepresent, independently, a lower alkyl or form together a saturatedhydrocarbon chain with 2 to 4 carbons:

a hydroxymethyl function is introduced into the general formula Fcompound, resulting in a general formula G compound ##STR15## in whichR₁, R₂₀, R₂₂, Z and Z' have the significance indicated above,

subsequently the alcoholic function of the compound of general formula Gis protected, resulting in the general formula H compound ##STR16## inwhich R₁, R₂₀, R₂₂, Z and Z' have the significance indicated above andR₂₃ represents a protector group of the alcohol group.

the acetal of the general formula H compound is unprotected, resultingin the general formula I' compound ##STR17## in which R₁, R₂₀, R₂₂, andR₂₃ have the significance indicated above,

the formula I' compound is treated with a functionalized alkylizingagent, resulting in a β-hydroxyester of general formula J ##STR18## inwhich R₁, R₂₀, R₂₂ and R₂₃ have the significance indicated above, R₁₈and R₁₉ are as defined in the general formula II and R₁₇ represents OR₆and R₆ represents a lower alkyl, cycloalkyl, lower cycloalkyl alkyl,lower alkenyl, lower alkoxy lower alkyl, or aryl or lower aryl alkyl;

the protector group R₂₃ is cleaved from the general formula J compound,resulting in a general formula K compound ##STR19## in which R₁, R₁₈,R₁₉, R₂₀ and R₂₂ have the significance indicated above and R₁₇represents OR₆ and R₆ represents a lower alkyl, cycloalkyl, lowercycloalkyl alkyl, lower alkenyl, lower alkoxy lower alkyl, or aryl orlower aryl alkyl;

general formula K compound is cyclized to form general formula Lcompound ##STR20## in which R₁, R₁₈, R₁₉, R₂₀ and R₂₂ have thesignificance indicated above, and finally

radical R₂₂ of the L compound is transformed into carbonyl, resulting inthe general formula M compound ##STR21## in which R₁, R₁₈, R₁₉ and R₂₀have the significance indicated above.

The carbonyl function of a 4-acyl-2-pyridine (obtained for exampleaccording to Lamattina, J. L., J. Heterocyclic Chem. 20, p. 553 (1983))is preferably shielded by an acetal function, preferably a cyclicacetal, according to the classic conditions known by persons skilled inthe art (Greene, T., Protective Groups in Organic Synthesis 10-86 (JohnWiley & Sons 1981)). When R22 is chloro or fluoro, the intermediary thusobtained is treated by a sodium alcoholate or potassium alcoholate in anaprotic solvent (for example acetonitrile), or in the alcohol from whichthe alcoholate is derived, at a temperature between 0° C. and 100° C.resulting in the general formula F compound. The latter may be lithiatedin position 3 by treatment with an aryl- or alkyl-lithium (for examplemesityl-lithium) in a ethered solvent such as tetrahydrofurane at atemperature between -100° C. and 0° C. A formylizing electrophile suchas N,N-dimethylformamide is added to the lithiated intermediary thusobtained, and the resulting aldehyde, after hydrolysis, is treated witha reducing agent such as sodium borohydride resulting in the generalformula G compound. The shielding of the alcohol function of compound Gis carried out according to classical conditions known by personsskilled in the art, resulting in a general formula H compound. Theexamples of the protector groups of the alcohol are comprised of thosethat form the ethers [for example methyl, methoxymethyl,tetrahydropyranyl, 2-methoxyethoxy methyl, benzyloxymethyl, t-butyl, andbenzyl (substituted or not)], and the esters (for example formate,acetate and isobutyrate). For other examples of other primary hydroxylprotector groups, refer to Greene, T., Protectives Groups in OrganicSynthesis. 10-86 (John Wiley & Sons, 1981). The unshielding of thegeneral formula H compounds resulting in the general formula I' compoundis performed in selective environments maintaining the integrity of theR₂₃ radical, for example, by treatment in acidic conditions (for examplewith trifluoroacetic acid). The selective conditions of protecting andunprotecting the functional groups are known by persons skilled in theart (Greene, T., Protective Groups in Organic Synthesis 10-86 (J. Wiley& Sons 1981)). The treatment of the I' compound by a functionalizedalkylizing agent resulting in a general formula J β-hydroxyester, may berealized with the help of a lithium enolate or a zinc derivative of acarboxylic ester in an anhydrous aprotic solvent, for exampletetrahydrofuran. The protector group R₂₃ of the general formula Jcompound, is cleaved resulting in the general formula K compound, underthe unprotected conditions known by persons skilled in the art. Forexample, when R₂₃ is a benzyl group, an alcoholic solution of generalformula J compound plus a palladium catalyzer may be subjected to ahydrogen atmosphere of 0.5 to 10 bars. The cyclization of the generalformula K compound thus obtained may be brought into acidic conditions(for example treatment with trifluoroacetic acid, or hydrochloric gasdissolved in an anhydrous solvent such as dichloromethane or dioxane)resulting in a β-hydoxylactonic cycle with seven links such as in thegeneral formula L compound. The general formula L compounds may betransformed into pyridones of general formula M, for example, by a hothydrochloric acid treatment, or with a trimethylsilyle iodide.

The 2-halo-3-quinoline-methanol of general formula N may be obtainedfrom the acetanilides of general formula P ##STR22## in which R₂, R₃ andR₄ have the significance indicated in the general formulas of compoundsI and II. In the procedures hereafter, the R₂, R₃ and R₄ groups may beprotected if necessary according to the classical methods of protection(Greene, T., Protective Groups in Organic Synthesis 10-86 (John Wiley &Sons 1981)).

Formula N compounds may then be obtained according to the followingprocedure: said anilines of formula P are N-acetylated by treatment withan acetylizing agent such as for example acetic anhydride. Theacetanilides thus obtained are treated at a temperature between 50° C.and 100° C., preferably 75° C., by a reagent known by persons skilled inthe art under the reagent name Vilsmeyer (obtained by the action ofphosphoryl oxychloride on N,N-dimethylformamide at a temperature between0° C. and 10° C.) resulting in corresponding2-chloro-3-quinolinecarbaldehyde (refer to, for example Meth-Cohn. etal., J. Chem. Soc., Perkin Trans. 1 p. 1520 (1981); Meth-Cohn, et al.,J. Chem. Soc., Perkin Trans. 1 p. 2509 (1981); and Nakasimhan et al., J.Am. Chem. Soc., 112. p. 4431 (1990)). The chlorine on position 2 of the2-chloro-3-quinolinecarbaldehydes may be substituted by iodine or bybromine by heating the product in an inert solvent such as acetonitrilein the presence of an iodine or bromine salt (for example sodium iodideor tetrabutylammonium bromide). A trace of acid such as concentratedhydrochloric acid may be necessary to catalyze this transformation. The2-halo-3-quinolinecarbaldehydes are easily reduced to2-halo-3-quinolinemethanols corresponding to the general formula N,under the classical conditions known by persons skilled in the art suchas the treatment in an alcoholic solvent (for example methanol) bysodium borohydride at a temperature between 0° C. and 40° C.

Formula N compounds may be obtained as well according to the followingprocedure: the general formula P anilines such as are defined above areacylated by reacting with a nitrile (such as chloroacetonitrile orpropionitrile) in the presence of boron trichloride and another Lewisacid such as aluminum trichloride, titanium tetrachloride ordiethylaluminum chloride in an aprotic solvent or a mixture of anaprotic solvent, followed by a hydrolysis (cf. Sugasawa, T., et al., J.Am. Chem. Soc. 100, p. 4842 (1978)). The resulting intermediate is thentreated with ethylmalonyl chloride in an aprotic solvent such asacetonitrile in the presence of a base such as triethylamine, thentreated by alkaline alcoholate, for example sodium methylate inmethanol, resulting in an ethyl 2-hydroxy-3-quinolinecarboxylatesubstituted in position 4. The latter is transformed into ethyl2-chloro-3-quinolinecarboxylate by treatment with aphosphoryloxychloride. When position 4 of the quinoline carries achloromethyl group, a nucleophilic substitution may be carried out bytreatment with a secondary amine such as for example dimethylamine,N-methylpiperazine, morpholine or piperidine. Ethyl2-chloro-3-quinolinecarboxylate is then reduced by diisobutylaluminiumhydride in an aprotic solvent such as dichloromethane resulting in2-chloro-3-quinolinemethanol of general formula N. The analogues of theintermediary (N) compounds were described in the literature and inparticular PCT Application 95/05427.

The invention has as well as its subject matter new industrial products,and notably as new industrial products destined for the production offormula I and II products, the formula I' and M products as is describedabove.

Certain compounds of the invention may be prepared in the form ofpharmaceutically acceptable salts according to the usual methods. Theacceptable salts comprise as an example and in a non-limiting manner,the addition salts of inorganic acids such as chlorohydrate, sulfate,phosphate, diphosphate, bromohydrate and nitrate or organic acids suchas acetate, maleate, fumarate, tartrate, succinate, citrate, lactate,methane sulfonate, p-toluenesulfonate, pamoate, salicylate, oxalate andstearate. The salts formed from bases such as sodium or potassiumhydroxide that enter as well into the field of application of thepresent invention when they are usable. For other examples ofpharmaceutically acceptable salts, refer to "Pharmaceutical Salts", J.Pharm. Sci. 66: 1 (1977).

The compounds of the present invention possess interestingpharmacological properties. This is the way in which the compounds ofthe present invention have an inhibiting effect on the topoisomerase Iand/or II and anti-tumor activity. The present technique suggests thatthe compounds of the invention present an antiparasitic and/or antiviralactivity. The compounds of the present invention may thus be used indifferent therapeutic applications.

The compounds may inhibit topoisomerase, for example of type I and/orII, in a patient, for example a mammal such as a human, byadministration to this patient of an effective therapeutic quantity of aformula (I) or a formula (II) compound.

The compounds of the invention possess as well anti-tumor activity. Theymay be used for the treatment of tumors, for example tumors expressing atopoisomerase, in a patient by administration to the former of aneffective therapeutic quantity of formula (I) or formula (II) compound.Some examples of tumors or cancers comprise cancers of the esophagus,the stomach, the intestines, the rectum, the oral cavity, the pharynx,the larynx, the lungs, the colon, the breast, the uterus cervix, theendometrium corpus, the ovaries, the prostate, the testicles, thebladder, the kidneys, the liver, the pancreas, the bones, the connectivetissue, the skin, the eyes, the brain and the central nervous system, aswell as cancer of the thyroid, leukemia, Hodgkin's disease, lymphomasother than Hodgkin's, multiple melanomas and others.

They may be used as well for the treatment of parasitic infections byinhibition of the hemoflagellates (for example in trypanosoma orleishmania infections) or by the inhibition of plasmodium (as forexample in malaria), but also the treatment of viral infections orillnesses.

These properties make formula I and II products suitable for apharmaceutical use. The present application has as well as its subjectmatter, as medications, the formula I and II products as is definedabove, as well as the addition salts with the pharmaceuticallyacceptable organic or mineral acids, said formula I and II products, aswell as the pharmaceutical compounds comprising, as an activeingredient, one of the medications as defined above.

The invention concerns thus pharmaceutical compositions comprising acompound of the invention or an additive salt of a pharmaceuticallyacceptable acid of it, in association with a pharmaceutically acceptablesupport following the chosen method of administration (for example oral,intravenous, intraperitoneal, intramuscular, transdermal orsubcutaneous). The pharmaceutical composition (for example therapeutic)may be in the form of a solid, liquid, liposome or lipid micell.

The pharmaceutical composition may be in the form of solids, forexample, powders, pills, granules, tablets, liposomes, capsules orsuppositories. The pill, tablet or capsule may be coated in a substancecapable of protecting the composition from the action of gastric acid orenzymes in the stomach of the subject during a sufficient period of timeto permit the composition to pass, non-digested into the small intestineof the latter. The compound may thus be administered locally, forexample by placement next to the tumor. The compound may also beadministered according to the time released method (for example a timereleased composition or an infusion pump). The solid appropriate mediummay be, for example, calcium phosphate, magnesium stearate, magnesiumcarbonate, talc, sugars, lactose, dextrine, amidon, gelatin, cellulose,methyl cellulose, sodium carboxymethyl cellulose, polyvinylpyrolidine,and wax. The pharmaceutical compositions containing a compound of theinvention may be presented as well in the form of a liquid like, forexample, solutions, emulsions, suspensions or a time-releaseformulation. The appropriate liquid supports may be, for example, water,organic solvents such as glycerol, or the glycols such as polyethyleneglycol, as well as their mixtures, in various proportions, in water.

The invention has as well as its subject matter the use of the formula Iand II products as is defined above, for the preparation of medicationintended to inhibit topoisomerase, and more particularly type I and typeII topoisomerase, of medication intended to treat tumors, of medicationsintended to treat parasitic infections, as well as medications intendedto treat viral infections or illnesses.

The dose of a compound according to the present invention, isanticipated for the treatment of illnesses or disorders mentioned above,vary according to the method of administration, the age and the bodyweight of the subject under treatment, as well as the condition of thelatter, and it will be decided definitively by the treating doctor orveterinarian is called here "therapeutically effective quantity".

In the following experimental section, an illustration of thepharmacological properties of the compounds can be found.

In the case that all the technical and scientific terms used here arenot defined in another manner, they have the same significance as theone currently understood by an ordinary specialist in the domain towhich the invention pertains. In the same way, all the publications,patent applications, all patents and all other references mentioned hereare incorporated by reference.

The following references are presented to illustrate the aboveprocedures and must not in any case be considered as limiting the scopeof the invention.

EXPERIMENTAL SECTION EXAMPLE 1 tert-butylβ-ethyl-β-hydroxy-γ-(8-hydroxymethyl-9-oxo(11H)-indolizino-[1,2-b]quinoline-7-yl)-propionate 1.a. 4-ethyl-3,4-dihydroxy-1H-pyrano [3',4':6,7] indolizino [1,2-b] quinoline-14 (4H,12H)-one

Sodium borohydride (14 g, 370 mmol) is added in portions to a suspensionof (S)-(+)-camptothecin (14 g, 40 mmol, that may be obtained usingdifferent commercial sources such as Aldrich Chemical Co. (Milwaukee,Wis.)), in methanol (750 ml) and the resulting mixture is heated slowlyto 55° C. so as to obtain a clear solution that is then agitated for 16hours at ambient temperature. The solvent is then evaporated underreduced pressure, the residue is recovered in water (250 ml),neutralized by the addition of acetic acid (21 ml) and left alone for 2hours at 4° C. The resulting suspension is filtered and washedsuccessively with cold water, acetone and diethyl ether, which, afterdrying under reduced pressure, permits obtaining the sought compound inthe form of a white solid, m.p. 280° C.

1.b. 8-formyloxymethyl-7-propionylindolizino [1,2-b] quinoline-9(11H)-one

A solution of sodium Metaperiodate (14 g, 65 mmol) in water (140 ml) isadded drop-wise to a suspension of 4-ethyl-3,4-dihydroxy-1H-pyrano[3',4': 6,7] indolizino [1,2-b] quinoline-14(4H,12H)-one (13.4 g, 38mmol) in glacial acetic acid (720 ml) and the resulting solution isagitated for one hour at ambient temperature. The reaction mixture isthen poured into a mixture of ice/water (650 ml) and the resultingsuspension is agitated for one half-hour then filtered and washedsuccessively with water, isopropyl alcohol, and etherdiethyl alcohol,which, after drying under reduced pressure, the sought compound (11.5 g)in the form of a pale yellow solid, m.p. >200° C. (d).

1.c. tert-butyl β-ethyl-β-hydroxy-γ-(8-hydroxymethyl-9-oxo(11H)-indolizino-[1,2-b] quinoline-7-yl)-propionate

A zinc suspension (6.5 g, 100 mmol) agitated with a magnetic agitator inanhydrous diethylether (50 ml) under argon, is activated, by drop-wiseaddition of chlorotrimethylsilane (0.75 ml, 5.7 mmol). Agitate again 15minutes at ambient temperature to heat to reflux. The heating bath isthen removed and tert-butyl bromoacetate (15 ml, 100 mmol) is addeddrop-wise at a rate assuring the maintenance of reflux. The externalheating is replaced and pursued again for one hour. The ethered solutionresulting from the Reformatsky reagent is left to cool to ambienttemperature then transferred, by means of a cannula, into a suspensionof 8-formyloxymethyl-7-propionylindolizino [1,2-b] quinoline-9 (11H)-one(1.6 g, 4.7 mmol) in anhydrous tetrahydrofuran (40 ml) under argon. Thereaction mixture is agitated at reflux for one hour, then left to coolto ambient temperature, and the reaction is stopped by adding saturatedammonium chloride (100 ml) and is extracted with chloroform (3×100 ml).The combined chloroformic extracts are dried on sodium sulfate,evaporated, and the residue is purified by gel chromatography on silicagel (1-2% MEOH/CH₂ Cl₂), which makes possible obtaining 0.64 g of thesought-after compound (31%) in the form of a pale yellow solid, m.p.146-149° C.

NMR-¹ H (CDCl₃); 0.93 (t, 3H); 1.37 (s, 9H); 1.99 (m, 2H); 2.97 (dd,2H); 3.5 (se, 1H); 5.10 (s, 2H); 5.24 (s, 2H); 7.40 (s, 1H); 7.59 (t,1H); 7.83 (t, 1H); 7.90 (d, 1H); 8.20 (d, 1H); 8.34 (s, 1H).

NMR-C¹³ (CDCl₃); 8.18; 27.90; 34.59; 45.34; 49.91; 58.55; 77.39; 82.42;100.52; 127.67; 127.97; 128.10; 128.64; 129.44; 129.79; 130.42; 130.99;142.86; 148.69; 152.75; 155.16; 162.38; 172.24.

IR (KBr); 764; 1016; 1157; 1580; 1651; 1726.

EXAMPLE 2 ethyl β-ethyl-β-hydroxy-γ-(8-hydroxymethyl-9 oxo(11H-indolizino-[1,2-b] quinoline-7-yl)-propionate

A suspension of zinc (500 mg, 7.64 mmol) and of8-formyloxymethyl-7-propionylindolizino [1,2-b] quinoline-9 (11H)-one(400 mg, 1.15 mmol) in anhydrous tetrahydrofuran (20 ml) containing 10mg of hydroquinone is heated to reflux under argon. The heating bath isremoved and the exothermic reaction initiated by the addition of a dropof ethyl bromoacetate and a small crystal of iodine. Reflux ismaintained by the drop-wise addition of ethyl bromoacetate (500 ml, 4.48mmol) then the reaction mixture is again heated at reflux for one hour.After cooling to ambient temperature, the reaction is stopped by addingsaturated ammonium chloride (10 ml) and methanol (30 ml). The resultantmixture is agitated for 5 minutes then filtered and evaporated. Theresidue is dissolved in dichloromethane (30 ml), washed with water anddried on sodium sulfate. This is followed by the elimination of thesolvent and a purification by column chromatography (SiO₂, CH₂ Cl₂ /MeOH98/2) resulting in 230 mg (49%) of the sought compound in the form of ayellow solid, m.p. 157-161° C.

NMR ¹ H (CDCl₃): 0.93 (t, 3H); 1.20 (t, 3H); 2.02 (m, 2H); 3.07 (dd,2H); 4.11(q, 2H); 4.9 (se, 1H); 5.08 (s, 2H); 5.23 (s, 2H); 7.45 (s,1H); 7.62 (t, 1H); 7.80 (t, 1H); 7.90 (d, 1H); 8.22 (d, 1H); 8.36 (s,1H).

NMR C¹³ (CDCl₃): 8.09; 14.01; 34.67; 44.85; 49.94; 58.31; 61.09; 77.21;100.78; 127.78; 127.96; 128.11; 128.72; 129.16; 129.65; 130.60; 131.32;142.76; 148.28; 152.55; 155.09; 162.22; 172.59.

IR (KBr): 766; 1009; 1184; 1582; 1647; 1750.

EXAMPLE 3 5-ethyl-4,5-dihydro-5-hydroxy-1H-oxepino [3',4':6,7]-indolizine [1,2-b]quinoline-3,15 (4H, 13H)-dione

tert-butyl β-ethyl-β-hydroxy-γ-(8-hydroxymethyl-9-oxo(11H)-indolizino-[1,2-b] quinoline-7-yl) propionate (1.45 g, 3.32 mmol)is dissolved in anhydrous dichloromethane (25 ml) and treated with asaturated solution of hydrogen chloride in dichloromethane (100 ml). Theresultant mixture is maintained at -20° C. for 16 hours. The precipitateis filtered, washed with methanol and dried under reduced pressure,which obtains 662 mg (55%) of the sought compound in the form of ayellow solid, m.p. >300° C.

NMR ¹ H (DMSO): 0.90 (t, 3H); 1.20 (q, 2H); 3.27 (dd, 2H); 5.29 (s, 2H);5.49 (dd, 2H); 7.42 (s, 1H); 7.73 (t, 1H); 7.90 (t, 1H); 8.16 (t, 2H);8.71 (s, 1H).

NMR C¹³ (DMSO): 8.45; 36.48; 42.54; 50.68; 61.44; 73.34; 99.78; 122.71;127.83; 128.15; 128.75; 129.08; 130.07; 130.61; 131.81; 144.66; 148.04;152.80; 155.91; 159.26; 172.08.

IR (KBr): 761; 1127; 1204; 1285; 1580; 1653; 1757.

EXAMPLE 4 β-ethyl-β-hydroxy-γ-(8-hydroxymethyl-9-oxo(11H)-indolizino-[1,2-b] quinoline-7-yl)-propionic acid

An aqueous solution of potassium hydroxide (0.1N, 30 ml) is added to5-ethyl-4,5-dihydro-5-hydroxy-1H-oxepino [3'4': 6,7]-indolizino [1,2-b]quinoline-3,15 (4H, 13H)-dione (500 mg, 1.38 mmol) and the resultantsuspension is agitated for 16 hours, which results in an almost clearliquid that is filtered. The filtrate is acidified to pH 3.5 with 1Nhydrochloric acid, and the yellow precipitate is recovered byfiltration, washed with water and acetone, then dried under reducedpressure. 415 mg (79%) of the sought compound is obtained in the form ofa monohydrate, m.p. 165-167° C.

NMR ¹ H (DMSO): 0.82 (t, 3H); 2.10 (m, 2H); 2.83 (d, 2H); 3.12 (d, 2H);3.25 (se, 1H); 4.81 (s,2H); 5.26 (s,2H); 5.76 (se, 1H); 7.38 (s, 1H);7.71 (t, 1H); 7.84 (t, 1H); 8.10 (d, 1H); 8.18 (d, 1H); 8.34 (s, 1H);12.15 (se, 1H).

NMR C¹³ (DMSO): 8.16; 34.80; 46.71; 50.36; 55.73; 76.53; 100.17; 127.50;128.00; 128.26; 128.69; 129.06; 130.01; 130.45; 131.63; 142.57; 148.09;153.19; 156.07; 161.22; 172.27.

IR (KBr): 1020; 1188; 1413; 1586; 1651; 1694.

EXAMPLE 5 methyl β-ethyl-β-hydroxy-γ-(8-hydroxymethyl-9-oxo(11H)-indolizino-[1,2-b] quinoline-7-yl)-propionate

5-ethyl-4,5-dihydro-5-hydroxy-1H-oxepino [3'4': 6,7]-indolizino [1,2-b]quinoline-3,15 (4H,13H)-dione (180 mg, 0.5 mmol), in suspension inmethanol (50 ml) is treated with 6N dry hydrogen chloride in methanol(0.5 ml) and maintained at reflux until completely dissolved (4 hours).The volatile compounds are evaporated and the residue is dissolved indichloromethane (50 ml), washed with diluted sodium hydroxide (0,05N, 15ml) and saline (15 ml). The organic fraction is dried on sodium sulfateand evaporated. The residual solid is purified by silica gel columnchromatography (MeOH at 3% / CH2Cl2) and the product is purified andrecovered in diethyl ether, filtered and dried, which gives 120 mg (58%)of the sought after compound in the form of a pale yellow solid, m.p.163-166° C.

NMR ¹ H (CDCl₃): 0.93 (t, 3H); 2.2 (m, 2H); 3.05 (dd, 2H); 3.49 (s, 3H);3,62 (s, 3H); 4.93 (s, 2H); 5.22 (d, 2H); 5.52 (s, 1H); 7.21 (s, 1H);7,62 (t, 1H); 7.81(t, 1H); 7.91 (d, 1H); 8.22 (d, 1H); 8.36 (s, 1H).

NMR C¹³ (CDCl₃): 7.74; 35.54; 46.82; 50.15; 51.67; 58.10; 65.33; 78.03;100.17; 125.57; 127.70; 128.04; 128.10; 128.35; 129.53; 130.39; 130.94;143.87; 148.75; 152.94; 157.83; 161.74; 171.35.

IR (KBr): 1207; 1595; 2655; 1709.

EXAMPLE 6 ethyl β-ethyl-α,α-difluoro-β-hydroxy-γ-(8-hydroxymethyl-9-oxo(11H)-indolizino-[1,2-b] quinoline-7-yl)-propionate

About one half of the total quantity of ethyl bromodifluoroacetate (1.8ml, 14 mmol), 8-formyloxymethyl-7-propionylindolizino [1,2-b]quinoline-9 (11H)-one (2.0 g, 5.75 mmol) such as is obtained in example1.b.) in suspension with anhydrous THF (10 ml), is added drop-wise underargon to a suspension of zinc (1.25 g, 17.2 mmol) in anhydrous THF atreflux (40 ml), then the remaining part of the ethylbromodifluoroacetate [is added]. The reaction mixture is maintained atreflux for another half-hour. After cooling to ambient temperature, thereaction is stopped by adding saturated aqueous ammonium chloride (20ml) and the reaction mixture is extracted with dichloromethane (3×20ml). The combined organic extracts are dried and concentrated. Theresidue is recovered in diethyl ether (10 ml), filtered and purified bycolumn chromatography (SiO₂, CH₂ Cl₂ /MeOH 98/2), resulting in 664 mg(26%) of the sought compound in the form of a yellow solid, m.p.208-209° C.

NMR ¹ H (CDCl₃): 0.91(t, 3H); 1.38 (t, 3H); 2.32 (m, 2H); 4.8 (se, 1H);4.38 (q. 2H); 5.09 (d, 2H); 5.13 (dd, 2H); 7.42 (s,1H); 7.55 (t, 1H);7.72 (t, 1H); 7.79 (d, 1H); 8.08 (d, 1H); 8.22 (s, 1H).

NMR C¹³ (CDCl₃): 6.97; 13.93; 28.63; 50.18; 56.27; 63.15; 77.20; 81.96(t); 101.27; 116.40 (t); 127.67; 127.77; 127.97; 128.31; 129.26; 130.33;130.94; 131.23; 143.16; 148.34; 150.20; 151.91; 161.21; 163.21 (t).

IR (KBr): 1124; 1308; 1591; 1647; 1748.

EXAMPLE 7 ethyl β-ethyl-β-hydroxy-γ-(8-hydroxymethyl-9-oxo(11H)-indolizino-[1,2-b] quinoline-7-yl)-propionate

A suspension of zinc (1.25 g, 19.1 mmol) of8-methyl-7-propionylindolizino [1,2-b] quinoline-9-(11H)-one (500 mg,1.43 mmol, such as is obtained by Kingsbury, W. D., Tetrahedron Lett.29: 6847 (1988)) and silver acetate (250 mg, 1.50 mmol) intetrahydrofuran (10 ml) is agitated at ambient temperature in an argonatmosphere. At the end of 10 minutes, the reaction mixture is activatedby the drop-wise addition of a molar solution of chlorodiethylaluminum(10 ml, 10 mmol), then ethyl bromoacetate (1.25 ml, 11.3 mmol) is addeddrop-wise and the resultant mixture is left to react for another 5hours. The reaction is stopped by the successive addition of ethylalcohol (10 ml) and a saturated solution of potassium and sodiumtartrate (10 ml). The resultant mixture is agitated again for one hour,filtered and concentrated under reduced pressure. The residue isrecovered in dichloromethane (30 ml), washed with water, dried,concentrated and purified by column chromatography (SiO₂, CH₂ Cl₂ /MeOH98/2), resulting in 93 mg (15%) of the sought product in the form of apale yellow solid, m.p. 185-188° C.

NMR ¹ H (CDCl₃): 0.91 (t, 3H); 1.17 (t, 3H); 1.99 (m, 2H); 2.49 (s, 3H);3.10 (dd, 2H); 4.11 (q, 2H); 4.6 (se, 1H) 5.25 (s, 2H); 7.65 (t, 1H);7.67 (s,1H); 7.80 (t, 1H); 7.90 (d, 1H); 8.22 (d, 1H); 8.34 (s, 1H).

NMR C¹³ (CDCl₃): 8.02; 13.99; 14.72; 33.14; 43.97; 50.02; 61.0; 76.54;101.90; 127.65; 127.84; 128.08; 128.81; 128.88; 130.74; 131.59; 131.65;140.33; 147.64; 152.96; 153.61; 162.11; 172.91.

IR (KBr): 762; 1192; 1576; 1653; 1740.

EXAMPLE 8 tert-butyl β-ethyl-β-hydroxy-γ-(8-hydroxymethyl-9-oxo(11H)-indolizino-[1,2-b] quinoline-7-yl)-propionate

Acetic anhydride (70 μl, 0.7 mmol) is added dropwise to a solution oftert-butyl β-ethyl-β-hydroxy-γ-(8-hydroxyinethyl-9-oxo(11H)-indolizino-[1,2-b] quinoline-7-yl)-propionate (200 mg, 0.46 mmol)and triethylamine (140 μl, 1 mmol) in dichloromethane (5 ml) and theresultant mixture is agitated at ambient temperature for 21 hours. Thevolatile compounds are evaporated and the residue is purified by asilica gel column chromatography (1-2% MeOH/CH₂ Cl₂), resulting in 152mg of the sought compound in the form of a yellow solid, m.p. 195-196°C.

NMR ¹ H (CDCl₃): 0.88 (t, 3H); 1.32 (s, 9H); 1.93 (m, 2H); 2.07 (s, 3H);2.97 (dd, 2H); 4.8 (se, 1H); 5.28 (s, 2H); 5.59 (dd, 2H); 7.39 (s, 1H);7.63 (t, 1H); 7.80 (t, 1H); 7.90 (d, 1H); 8.23 (d, 1H); 8.34 (s, 1H).

NMR C¹³ (CDCl₃): 8.02; 21.06; 27.91; 35.05; 45.58; 50.16; 59.23; 77.52;82.26; 100.59; 124.21; 127.91; 128.10; 128.14; 128.97; 129.18; 130.68;131.46; 142.85; 148.29; 152.43; 158.49; 161.83; 171.13; 171.90.

EXAMPLE 9 5,12-diethyl-4,5-dihydro-5-hydroxy-1H-oxepino [3',4':6,7]-indolizino [1,2-b] quinoline-3,15 (4H,13H)-dione

This compound is prepared in a manner analogous to example 1, exceptthat in stage 1.a. the 7-ethyl camptothecin (Sawada et al., Chem. Pharm.Bull. 39: 2574 (1991)) is used in place of the camptothecin. The soughtcompound is obtained in the form of a bright yellow solid, m.p. >270° C.

NMR ¹ H (DMSO): 0.92 (t, 3H); 1.39 (t, 3H); 1.93 (q, 2H); 3.08 (d, 2H);3.25 (q, 2H); 3.51 (d, 2H); 5.32 (s, 2H); 5.52 (dd, 2H); 7.42 (s, 1H);7.76 (t, 1H); 7.89 (t, 1H); 8.18 (d, 1H); 8.32 (d, 1H).

NMR C¹³ (DMSO): 8.46; 14.15; 22.42; 36.50; 42.54; 49.95; 61.45; 73.35;99.68; 122.61; 124.27; 126.76; 127.70; 128.27; 129.92; 130.18; 145.17;145.82; 148.57; 152.15; 155.89; 159.26; 172.08.

EXAMPLE 10 β-ethyl-γ-(12-ethyl-8-hydroxymethyl-9-oxo(11H)-indolizino-[1,2-b] quinoline-7-yl)-β-hydroxy-propionic acid

This compound is prepared in a manner analogous to example 4, exceptthat 5,12-diethyl-4,5-dihydro-5-hydroxy-1H-oxepino [3'4':6,7]-indolizino [1,2-b] quinoline-3,15 (4H,13H)-dione is used in placeof 5-ethyl-4,5-dihydro-5-hydroxy-1H-oxepino [3'4': 6,7]-indolizino[1,2-b] quinoline-3,15 (4H,13H)-dione. It is obtained in the form of aslightly off-white solid, m.p. 238-239° C.

NMR ¹ H (DMSO): 0.82 (t, 3H); 1.35 (t, 3H); 2.01 (m, 2H); 2.85 (d, 2H);3.18 (d, 2H); 3.22 (q, 2H); 4.81 (s, 2H); 5.00 (se, 1H); 5.24 (s, 2H);5.78 (se, 1H); 7.38 (s, 1H); 7.77 (t, 1H); 7.86 (t, 1H); 8.18 (d, 1H);8.28 (d, 1H); 12.10 (se, 1H).

NMR C¹³ (DMSO): 8.12; 14.15; 22.41; 34.78; 46.74; 49.65; 55.71; 76.51;100.04; 124.22; 126.63; 127.48; 128.12; 128.21; 129.94; 130.02; 143.10;145.59; 148.69; 152.62; 156.03; 161.22; 172.22.

EXAMPLE 11 8-ethyl-2,3,8,9-tetrahydro-8-hydroxy-10H-[1,4]dioxino [2,3-g]oxepino[3',4':6,7] indolizino [1,2-b] quinoline-10,13 (15H)-dione 11.a.2-ethyl-2-(2-methoxy-4-pyridyl)-1,3dioxolane (F)

Azeotropic water is distilled for one entire night with a Dean Starkapparatus from a mixture of 2-chloro-4-propionylpyridine (10 g, 59 mmol)obtained as in Lamattina, J. L., J. Heterocyclic Chem. 20, p. 553(1983)), of ethylene glycol, and of p-toluenesulfolic acid 250 mg) intoluene 150 ml). The solvent is then eliminated under reduced pressure,the acid is neutralized with saturated aqueous sodium bicarbonate (100ml) and the product is extracted using ether. The combined etheredextracts are washed with saline, dried on sodium sulfate and evaporated,which produces 13.3 g (96%) of the crude product protected by thecarbonyl group which is brought to reflux with 3 equivalents of sodiummethoxide in acetonitrile until the end of the reaction (control by thinbed chromatography: SiO₂, tert-butyl methyl oxide / hexane (TBMO/HX)50/50). The acetonitrile solution is then filtered and evaporated. Theresidue is recovered in ether, washed with water and saline, dried onsodium sulfate and evaporated, resulting in a brown oil which isdistilled (70-75° C, 0.04 mbar); 10.7 g (global return 81%) of product(F) is recovered in the form of a clear liquid.

11.b. 2-ethyl-2-(3-hydroxymethyl-2-methoxy-4-pyridyl)-1,3-dioxolane (G)

Tert-butyllithium (1.7 M in pentane, 100 ml, 170 mmol) is addeddrop-wise using a cannula to a solution of bromomesitylene (13 ml, 85mmol) in anhydrous tetrahydrofurane (300 ml) at -78° C. under argon. Theresultant white precipitate is agitated at -78° C. for one hour then2-ethyl-2-(2-methoxy-4-pyridyl)-1,3-dioxolane (10 g, 44.8 mmol) is addedand the reaction mixture is agitated 15 minutes at -78° C., one hour at0° C. and one hour at ambient temperature. After another cooling to -78°C., anhydrous N,N-dimethylformamide (100 mmol) is added and the reactionmixture is left to reheat to ambient temperature then is agitated for 16hours, after which there is an analysis by thin bed chromatography(SiO₂, TBMO/HX: 50/50) makes the complete consumption of the startingcompound. The reaction is stopped with saturated ammonium chloride andthe reaction mixture is extracted with diethyl ether (200 ml, 50 ml, 50ml). The combined extracts are dried on sodium sulfate and evaporated,resulting in a yellow oil which is purified by column chromatography(SiO₂, TBMO/HX: 0/100 to 5/95 in order to elute the mesytylenederivatives then 20/80 to 50/50 to elute the product) resulting in theintermediary aldehyde (7 g). The aldehyde is dissolved in methanol (100ml) and treated with sodium borohydride (5 g, 132 mmol) and theresultant mixture is agitated until the complete consumption of theintermediary aldehyde (about 1 hour) by analytic determination by thinbed chromatography. The solvent is then evaporated, the residue isrecovered in ether, washed with water and saline, dried, and the solventis evaporated. The column chromatography (SiO₂, TBMO/HX: 10/90 to 50/50)of the residue gives 7 g (global return 62%) of product (G) in the formof a yellow oil.

11.c. 2-(3-benzyloxymethyl-2-methoxy-4-pyridyl)-2-ethyl-1,3-diocolane(H)

A solution of2-ethyl-2-(3-hydroxymethyl-2-methoxy-4-pyridyl)-1,3-dioxolane (7 g, 30mmol) and of benzyl chloride (5 ml, 45 mmol) in anhydroustetrahydrofuran (150 ml) is added drop-wise to a suspension of sodiumhydride (80% in mineral oil, 1.85 g, 61 mmol) in anhydroustetrahydrofuran (100 ml) and the reaction mixture is maintained atreflux for 16 hours. The reaction mixture is then left to cool toambient temperature, the reaction is stopped with water (50 ml) and thereaction mixture is concentrated under reduced pressure. The residue isdissolved in diethyl ether (150 ml) and washed with water and saline,dried and evaporated. A purification by column chromatography (SiO₂,TBMO/HX; 5/95 to 20/80) results in the product protected by the benzyl(H). 9 g. (87%) in the form of a clear oil.

11.d. 1-(3-benzyloxymethyl-2-methoxy-4-pyridyl)-propane-1-one (I')

2-(3-benzyloxymethyl-2-methoxy4-pyridyl)-2-ethyl-1,3-dioxolane (9 g, 27mmol) is treated with trifluoracetic acid (10 ml) and water (5 ml) in atemperature bath of 120° C. for 3 hours. The reaction mixture isconcentrated under reduced pressure and the residual traces of the acidsare neutralized by the addition of a saturated aqueous sodiumbicarbonate. An ether extraction is followed by column chromatography(SiO₂, TBMO/HX; 10/90) results in 5.5 g (70%) of product (I).

11.e. tert-butyl β-ethyl-β-hydroxy-γ-(3-benzyloxymethyl-2-methoxy-4pyridyl) propionate

Tert-butyl bromoacetate (13 ml, 80 mmol) is added drop-wise to asuspension of zinc (5.3 g, 80 mmol activated by treatment with 6N HClfor 10 seconds, then washed successively with water until neutral pH,acetone and diethyl ether) in anhydrous tetrahydrofuran (60 ml) toreflux. The reaction medium is maintained at reflux for another 10minutes after which the addition is finished. The a solution of1-(3-benzyloxymethyl-2-methoxy-4-pyridyl)propane-1-one (5.8 g, 20 mmol)in anhydrous tetrahydrofuran (20 ml) is added, and the reaction mixtureis agitated under reflux for one more hour. The reaction is stopped at0° C. with saturated aqueous ammonium chloride (100 ml) and the reactionmixture extracted with diethyl ether. The combined extracts are dried onsodium sulfate and evaporated, resulting in a yellow oil which ispurified by column chromatography (SiO₂, TBMO/HX: 5/95 to 10/90) toobtain the tert-butyl ester (j)(7 g 8 95%) in the form of a clear oil.

11.f. tert-butylβ-ethyl-β-hydroxy-γ-(3-hydroxymethyl-2-methoxy-4-pyridyl) propionate

Tert-butyl β-ethyl-β-hydroxy-γ-(3-hydroxymethyl-2-methoxy-4-pyridyl)propionate (1 g, 2.5 mmol) is hydrogenolised at atmospheric pressure andambient temperature by using 5% of palladium on charcoal as a catalyst(50 mg) and absolute ethanol as solvent (10 ml). Once the reaction isfinished (6 hours), the catalyst is separated by filtration and thesolvent is evaporated, which yields 0.7 g (90%) of product (K) of asufficient purity for a subsequent synthetic use.

11.g. 5-ethyl-1,5-dihydro-5-hydroxy-9-methoxy-oxepino [3,4-c] pyridine-3(4H)-one (L)

Tert-butyl β-ethyl-β-hydroxy-γ-(3-hydroxymethyl-2-methoxy-4-pyridyl)propionate (8.8 g, 28 mmol) is treated with trifluoroacetic acid (30 ml)for 3 hours at ambient temperature. The volatile components areevaporated and the residue is purified by column chromatography (SiO₂,CH₂ Cl₂ /MeOH: 100/0 to 98/2), which results in a clear oil, aftertreatment with toluene, gives 5.9 g of product (L) (89%) in the form ofwhite crystals, m.p. 97-98° C.

11.h. 5-ethyl-1,5-dihydro-5-hydroxy-oxepion [3,4-c] pyridine3,9(4H,8H)dione (M)

Hydrochloric acid 1N (20 ml),5-ethyl-1,5-dihydro-5-hydroxy-9-methoxy-oxepino [3,4-c] pyridine-3(4H)-one (0.5 g, 2.1 mmol) is heated to reflux for 9 hours. The reactionmixture is concentrated under pressure and the residue is again dried bythe addition and evaporation of toluene, two times, then left overnightunder reduced pressure in the presence of phosphorous pentoxide. Theresultant oil is dissolved in anhydrous acetonitrile (5 ml) and agitatedunder argon for 24 hours. The precipitate is filtered and dried, whichyields 0.23 g (49%) of a white solid (M), m.p. 118-119° C.

11.i. 6,7-ethylenedioxy-2-iodo-3-quinoline-methanol (N)

The procedures described by Meth-Cohn et al; J. Chem. Soc. Perkin Trans.I, p. 1520 (1981); Meth-Cohn, J. Chem. Soc. Perkin Trans. I, p. 2509(1981), and Nakasimhan et al, J. Am. Chem. Soc. 112, p. 4431 (1990), areused. 3,4-ethylenedioxyacetanilide (22 g, 113 mmol) is added to theVilsmeyer reagent obtained by the drop-wise addition of phosphoryloxychloride (71 ml, 0.77 mol) to anhydrous dimethylformamide (23 ml,0.28mol), chilled in an ice-water bath for another 0.5 hour in an argonatmosphere. The resultant mixture is heated to 75° C. for 16 hours.After cooling to ambient temperature, the reaction mixture is added to amixture of ice and water (300 ml) and extracted with dichloromethane(5×200 ml). The combined organic extracts are dried on sodium sulfate,filtered and concentrated. The solid residue is placed into suspensionin dichloromethane (20 ml), filtered and dried under reduced pressure,which results in 10 g (35%) of2-chloro-6,7-ethylenedioxyquinoline-3-carbaldehyde in the form of ayellow solid, m.p. 222-224° C. This intermediary is treated with sodiumiodide (30 g, 0.2 mol) and concentrated hydrochloric acid (1.5 ml) inacetonitrile to reflux (150 ml) for 24 hours. After cooling to ambienttemperature, the solvent is eliminated under reduce pressure and theresidue is recovered in 50% aqueous tetrahydrofuran (200 ml), filtered,washed with tetrahydrofuran and dried under reduced pressure, whichyields 12 g of 6,7-ethylenedioxy-2-iodoquinoline-3-carbaldehyde in theform of a yellow solid, m.p. 155-157° C. The above intermediary istreated with sodium borohydride (2 g, 52 mmol) in methanol (200 ml) atambient temperature for 0.5 hours. The solvent is eliminated underreduced pressure and the residue is recovered in water and filtered. Theresultant solid is dried under reduced pressure in the presence ofphosphorous pentoxide, resulting in 11 g of(6,7-ethylenedioxy-2-iodoquinoline-3-yl)-methanol in the form of ayellow solid, m.p. 178-180° C.

11.j.5-ethyl-8-(6,7-ethylenedioxy-2-iodo-3-quinolinemethyl)-1,5-dihydro-5-hydroxy-oxepino[3,4-c] pyridine 3,9(4H,8H)-dione (O)

Over 5 minutes, diethyl azodicarboxylate (570 μl, 3.6 mmol) is addeddrop-wise to a solution of 5-ethyl-1,5dihydro-5-hydroxy-oxepino [3,4-c]pyridine 3,9(4H,8H)-dione (400 mg, 179 mmol), of compound obtained inthe preceding stage 11.i. (770 mg, 2.23 mmol) and triphenylphosphine(934 mg, 3.58 mmol) in a mixture of anhydrous THF/DMSO (8/1 v/v, 45 ml)and the resultant mixture is agitated under argon at ambient temperaturefor 16 hours. The reaction mixture is then concentrated under reducedpressure and the residue is dissolved in chloroform (100 ml). Theresultant solution is washed with saline (4×50 ml), dried on sodiumsulfate and evaporated. The residue is purified by column chromatography(SiO₂, CH₂ Cl₂ / MEOH: 99/1 to 98/2), resulting in 650 mg (66%) ofproduct (O) in the form of a white solid, m.p. 165-167° C.

11.k. 8-ethyl-2,3,8,9-tetrahydro-8-hydroxy-10H,12H-[1,4] dioxino [2,3-g]oxepino [3',4':6,7] indolizino [1,2-b] quinoline-10,13 (15H )-dione

5-ethyl-8-(6,7-ethylenedioxy-2-iodoquinoline-3-yl)methyl-4,5-dihydro-5-hydroxy-(1H,3H)oxepino[3,4-c] pyridine-3-dione (600 mg, 1.1 mmol), tetrabutyl-ammonium bromide(352 mg, 1.1 mmol), sodium acetate (359 mg, 4.4 mmol) and palladium IIacetate (98 mg, 0.43 mmol) are dissolved in anhydrous acetonitrile (40ml) and heated to 90° C. under argon for 16 hours. After cooling toambient temperature, a white precipitate is separated from the reddishsolution. This precipitate is filtered and dried under reduced pressure.The crude product is put into suspension in water, filtered and driedunder reduced pressure on phosphorous pentoxide, resulting in 250 mg ofthe sought compound in the form of a clear yellow solid, m.p. >250° C.

NMR-¹ H (DMSO): 0.91 (t, 3H); 1.87 (m, 2H); 3.08 (d, 1H); 3.51 (d, 1H);4.45 (s, 2H); 5.19 (s, 2H); 5.47 (dd, 2H); 6.02 (se, 1H); 7.33 (s, 1H);7.54 (s, 1H); 7.55 (s, 1H); 8.43 (s, 1H).

NMR-C¹³ (DMSO): 8.43; 36.47; 42.54; 50.52; 61.43; 64.43 (2C); 73.31;99.07; 112.27; 113.14; 122.00; 124.24; 128.18; 129.74; 144.59; 145.01;145.33; 147.63; 150.88; 155.88; 159.23; 172.07.

EXAMPLE 12 10-benzyloxy-5-ethyl-4,5-dihydro-5-hydroxy-1H-oxepino[3',4':6,7]-indolizino [1,2-b] quinoline-3,15 (4H,13H)-dione 12.a.(6-benzyloxy-2-iodo-3-quinoline)-methanol

This compound is prepared in a method analagous to that outlined instage 11.i. of example 11, but in using 4-benzyloxyacetanilide in placeof 3,4-ethylene-dioxyacetanilide. A purification by columnchromatography on silica gel and the use of dichloromethane as eluantare necessary to isolate (8% return) to a sufficient purity theintermediary 6-benzyloxy-2-chloroquinoline-3-carbaldehyde, m.p. 180-182°C. Next, the exchange of the subsequent halogen reduction to sodiumborohydride results in (6-benzyloxy-2-iodoquinoline-3-yl)-methanol, m.p.147-149° C.

12.b.8-(6-benzyloxy-2-iodo-3-quinolinemethyl)-1,5-dihydroxy-5-ethyl-5-hydroxy-oxepino[3,4-c]pyridine-3,9(4H,8H)-dione

This compound is prepared in a method analagous to that outlined instage 11.j. of example 11, but using(6-benzyloxy-2-iodoquinoline-3-yl)-methanol in place of6,7-ethylenedioxy-2-iodoquinoline-3-yl)-methanol. This compound appearsin the form of a white solid, m.p. 197-199° C.

12.c. 10-benzyloxy-5-ethyl-4,5-dihydro-5-hydroxy-1H-oxepino[3',4':6,7]-indolizino [1,2-b] quinoline-3,15 (4H,13H)-dione

This compound is prepared in a method analagous to the one indicated instage 11.k. of example 11, but using8-(6-benzyloxy-2-iodo-3-quinolinemethy)-1,5-dihydroxy-5-ethyl-5-hydroxy-oxepino[3,4-c] pyridine-3,9(4H,8H)-dione in place of5-ethyl-8-(6,7-ethylenedioxy-2-iodoquinoline-3-yl)methyl-4,5-dihydro-5-hydroxy-(1H,3H)oxepino [3,4-c] pyridine-3-dione. The sought compound appears in theform of a clear yellow solid, m.p. >250° C.

NMR-¹ H (DMSO): 0.90 (t, 3H); 1.85 (m, 2H); 3.08 (d, 1H); 3.50 (d, 1H);5.25 (s, 2H); 5.30 (s, 2H); 5.50 (dd, 2H); 6.05 (s, 1H); 7.30-7.70 (m,8H); 8.10 (d, 1H); 8.55 (s, 1H).

NMR-C¹³ (DMSO): 8.43; 36.48; 38.28; 50.65; 61.42; 70.00; 73.32; 99.05;107.71; 122.05; 123.42; 128.18; 128.26; 128.70; 129.40; 130.19; 130.48;130.63; 136.65; 144.18; 144.90; 150.53; 155.91; 157.31; 159.24; 172.06.

EXAMPLE 13 γ-(12-benzyloxy-8-hydroxymethyl-9-oxo (11H)-indolizino[1,2-b] quinoline-7-yl)-β-ethyl-β-hydroxy-propionic acid (E)

This compound is prepared in a manner analagous to the one explained inexample 4, but using10-benzyloxy-5-ethyl-4,5-dihydro-5-hydroxy-1H-oxepino[3',4':6,7]-indolizino [1,2-b]quioline-3,15 (4H,13H)-dione in place of5-ethyl-4,5-dihydro-5-hydroxy-1H-oxepino [3',4':6,7]-indolizino [1,2-b]quinoline-3,15 (4H,13H)-dione. It appears in the form of a yellow solid,m.p. 171-173° C.

NMR-¹ H (DMSO): 0.80 (t, 3H); 2.00(m, 2H); 2.85 (d, 1H); 3.15(d, 1H);4.80 (s, 2H); 5.25 (s, 2H); 5.30 (s, 2H); 5.75 (se, 1H); 7.30 (s, 1H);7.35-7.70 (m, 7H); 8.10 (d, 1H); 8.55 (s, 1H).

NMR-C¹³ (DMSO): 8.11; 34.75; 46.68; 50.35; 55.70; 69.97; 76.51; 99.45;107.78; 123.28; 127.64; 128.18 (2C); 128.26; 128.70 (2C); 129.33;130.17; 130.47; 130.57; 136.69; 142.79; 144.17; 150.93; 156.03; 157.19;161.20.

EXAMPLE 14 5-ethyl-4,5-dihydro-5,10,-dihydroxy-1H-oxepino[3',4':6,7]-indolizino [1,2-b] quinoline-3,15 (4H,13H)-dione

10-benzyloxy-5-ethyl-4,5-dihydro-5-hydroxy-1H-oxepino[3',4':6,7]-indolizino [1,2-b] quioline-3,15 (4H,13HO-dione (370 mg,0.79 mmol) is treated with hydrogen at atmospheric pressure and atambient temperature by using 10% palladium on charcoal as catalyst (60mg) and trifluoroacetic acid as solvent (15 ml). Once the reaction isterminated (16 hours), dichloromethate (50 ml) and methanol (50 ml) areadded to the reaction mixture, the catalyst is filtered and the volatilecomponents are evaporated under reduced pressure, which permitsobtaining the crude form of the sought compound containing traces oftrifluoroacetic acid. These traces are eliminated by codistillation with1,4-dioxane. The product obtained is in the form of an orange solid,m.p. 150° C. (d), with a purity sufficient for any further syntheticuse.

NMR-¹ H (DMSO) 0.89 (t,3H); 1.85(q, 2H); 3.02 (d, 1H); 3.45 (d, 1H);5.19 (s, 2H); 5.37 (d, 1H); 5.50 (d, 1H); 5.98 (se, 1H); 7.26 (s, 1H);7.31 (s, 1H); 7.40 (d, 1H); 8.00 (d, 1H); 8.42 (s, 1H); 10.32 (s, 1H).

NMR-C¹³ (DMSO): 8.47; 36.50; 42.61; 50.57; 61.46; 73.35; 98.84; 109.02;121.83; 123.18; 129.50; 129.85; 130.12; 130.80; 143.39; 145.10; 149.69;155.97; 156.82; 159.30; 172.11.

EXAMPLE 1511-(dimethylamino)methyl-5-ethyl-4,5-dihydro-5,10-dihydroxy-1H-oxepino[3',4':6,7]-indolizino [1,2-b] quinoline-3,15 (4H,13H)-dione 15.a. 11-(dimethylarnino)methyl-5-ethyl-4,5-dihydro-5,10-dihydroxy-1H-oxepino[3',4':6,7]-indolizino [1,2-b] quinoline-3,15 (4H,13H)-dione

A suspension of 10-benzyloxy-5-ethyl-4,5-dihydro-5-hydroxy-1H-oxepino[3',4':6,7]-indolizino [1,2-b] quioline-3,15 (4H,13H)-dione (260 mg,0.69 mmol) in acetic acid (15 ml) is treated with 37% aqueousformaldehyde (500 μl) and 40% aqueous dimethylamine (500 μl) and theresultant mixture is agitated at ambient temperature for 16 hours. Thereaction mixture is concentrated to dryness and the residue is purifiedby a column chromatography (SiO₂, CH₂ Cl₂ /MeOH: 100/0 to 90/10)followed by crystallization with acetonitrile, resulting in 102 mg ofthe sought compound.

15.b.11-(dimethylamino)methyl-5-ethyl-4,5-dihydro-5,10-dihydroxy-1H-oxepino[3',4':6,7]-indolizino [1,2-b] quinoline-3,15 (4H,13H)-dionechlorhydrate

Diluted hydrochloric acid (1N) is added drop-wise to a suspension of11-dimethylamino)methyl-5-ethyl-4,5-dihydro-5,10-dihydroxy-1H-oxepino[3',4':6,7]-indolizino [1,2-b] quinoline-3,15 (4H,13H)-dione (102 mg) inwater, until completely dissolved. The water is evaporated under reducedpressure and the residue is put into suspension in acetonitrile (5 ml)and filtered, resulting in 103 mg of the sought salt, m.p. 248° C. (d).

NMR-¹ H (DMSO): 0.88 (t, 3H); 1.85 (m, 2H); 2.84 (s, 6H); 3.08 (d, 1H);3.5 (d, 1H); 4.73 (s, 2H); 5.47 (dd, 2H); 7.33 (s, 1H); 7.38 (s, 1H);7.72 (d, 1H); 8.19 (d, 1H); 8.99 (s, 1H); 9.92 (se, 1H); 11.45 (s, 1H).

NMR-C¹³ (DMSO): 8.46; 34.36; 42.44 (3C); 50.61 (2C); 61.42; 73.35;99.19; 108.63; 122.21; 122.36; 126.86; 129.13; 130.61; 133.09; 143.53;144.70; 149.76; 155.98; 157.17; 159.27; 172.06.

EXAMPLE 16 5-ethyl-9-fluoro-4,5-dihydro-5-hydroxy-10-methoxy-1H-oxepino[3',4':6,7] indolizino [1,2-b] quinoline-3,15 (4H,13H)-dione

This compound is obtained from 3-fluoro-4-methoxyaniline according tothe method illustrated by the stages 11.i, 11.j, and 11.k of example 11.Yellow solid, m.p. >250° C.

NMR-¹ H (DMSO): 0.89 (t, 3H); 1.85 (q, 2H); 3.08 (d, 1H); 3.49 (d, 1H);4.00 (s, 3H); 5.25 (s, 2H); 5.39 (d, 1H); 5.51 (d, 1H); 6.00 (s, 1H);7.32 (s, 1H); 7.72 (d, 1H); 7.91 (d, 1H); 8.58 (s,1H).

NMR-C¹³ (DMSO): 8.43; 36.48; 42.51; 50.68; 56.60; 61.42; 73.29; 99.25;108.68; 113.52; 122.23; 126.33; 129.99; 130.30; 143.79; 144.70; 148.42;151.18; 153.19; 155.81; 159.20; 172.06.

IR (KBr): 1259; 1503; 1602; 1737.

EXAMPLE 17 9-chloro-5-ethyl-4,5-dihydro-5-hydroxy-10-methyl-1H-oxepino[3',4':6,7] indolizino [1,2-b] quinoline-3,15 (4H,13H)-dione

This compound is obtained from 3-chloro-4-methoxyaniline according tothe method illustrated by the stages 11.i, 11.j, and 11.k of example 11.Yellow solid, m.p. >250° C.

NMR-¹ H (DMSO): 0.85 (t, 3H); 1.85 (q, 2H); 2.55 (s, 3H); 3.07 (d, 1H);3.45 (d, 1H); 5.25 (s, 2H); 5.39 (d, 1H); 5.51 (d, 1H); 6.05 (s, 1H);7.39 (s, 1H); 8.10(s, 1H); 8.20(s,1H); 8.60(s,1H).

NMR-C¹³ (DMSO): 8.43; 20.20; 36.47; 42.49; 50.67; 61,41; 73.28; 99.87;122.82; 126.98; 127.99; 129.60; 130.53; 131.08; 135.64; 136.56; 144.39;147.11; 153.10; 155.85; 159.18; 172.03.

IR (KBr): 1208; 1479; 1606; 1656; 1724.

EXAMPLE 18 5-ethyl-9,10-difluoro-4,5-dihydro-5-hydroxy-1H-oxepino[3',4':6,7] indolizino [1,2-b] quinoline-3,15 (4H,13H)-dione

This compound is obtained from 3,4-difluoroaniline according to themethod illustrated by the stages 11.i, 11.j, and 11.k of example 11.Yellow solid, m.p. >250° C.

NMR-¹ H (DMSO): 0.85 (t, 3H); 1.85 (q, 2H) ; 3.07 (d, 1H); 3.47 (d, 1H);5.25 (s, 2H); 5.39 (d, 1H); 5.51 (d, 1H); 6.05 (s, 1H); 7.39 (s, 1H);8.15(q, 1H); 8.25(q,1H); 8.68(q,1H).

NMR-C¹³ (DMSO): 8.41; 36.45; 42.48; 50.68; 61.40; 73.25; 99.92; 114.14;115.42; 115.58; 122.96; 125.52; 130.56; 131.46; 144.21; 145.25; 142.36;153.41; 155.85; 159.15; 172.00.

IR (KBr): 1266; 1512; 1581; 1618; 1751.

EXAMPLE 19 7-ethyl-7,8-dihydro-7-hydroxy-9H,11H-[1,3] dioxole [4,5-g]oxepino [3',4':6,7] indolizino [1,2-b] quinoline-9,12 (14H)-dione

This compound is obtained from 3,4-methylenedioxyaniline according tothe method illustrated by the stages 11.i, 11.j, and 11.k of example 11.Beige solid, m.p. >250° C.

NMR-¹ H (DMSO): 0.85 (t, 3H); 1.85 (q, 2H) ; 3.07 (d, 1H); 3.45 (d, 1H);5.20 (s, 2H); 5.39 (d, 1H); 5.51 (d, 1H); 6.00 (s, 1H); 6.30 (s, 1H);7.30(s, 1H); 7.49(d,2H); 8.45(s,1H).

NMR-C¹³ (DMSO): 8.43; 36.49; 42.56; 50.58; 61.42; 73.31; 98.87; 102.75;103.33; 104.92; 121.76; 125.74; 128.59; 130.33; 145.08; 146.69; 148.78;150.19; 151.49; 155.90; 159.24; 172.08.

IR (KBr): 1248; 1459; 1606; 1731.

EXAMPLE 20 9-chloro-5-ethyl-4,5-dihydro-5-hydroxy-10-methoxy-1H-oxepino[3',4':6,7] indolizino [1,2-b] quinoline-3,15 (4H,13H)-dione

This compound is obtained from 3-chloro-4-methoxyaniline according tothe method illustrated by the stages 11.i, 11.j, and 11.k of example 11.White solid, m.p. >250° C.

NMR-¹ H (DMSO): 0.85 (t, 3H); 1.85 (q, 2H) ; 3.07 (d, 1H); 3.45 (d, 1H);4.01 (s, 3H); 5.22 (s, 2H); 5.39 (d, 1H); 5.51 (d, 1H); 6.02 (s, 1H);7.31 (s, 1H); 7.68(s, 1H); 8.20(s, 1H); 8.55(s,1H).

NMR-C¹³ (DMSO): 8.82; 36.27; 42.30; 50.48; 56.69; 61.23; 73.08; 99.16;107.44; 122.16; 127.12; 128.12; 128.25; 130.02; 130.53; 143.29; 144.37;151.12; 153.29; 155.71; 158.98; 171.84.

IR (KBr): 1056; 1256; 1483; 1592; 1657; 1747.

EXAMPLE 21 5-ethyl-4,5-dihydro-5-hydroxy-10-methoxy-1H-oxepino[3',4':6,7] indolizino [1,2-b] quinoline-3,15 (4H,13H)-dione

This compound is obtained from 4-methoxyaniline according to the methodillustrated by the stages 11.i, 11.j, and 11.k of example 11. Yellowsolid, m.p. >250° C.

NMR-¹ H (DMSO): 0.85 (t, 3H); 1.85 (q, 2H) ; 3.07 (d, 1H); 3.45 (d, 1H);3.95 (s, 3H); 5.28 (s, 2H); 5.40 (d, 1H); 5.51 (d, 1H); 6.00 (s, 1H);7.38 (s, 1H); 7.51(d, 2H); 8.07(d,1H); 8.55(s,1H).

NMR-C¹³ (DMSO): 8.45; 36.48; 42.51; 50.64; 55.92; 61.42; 73.33; 99.01;106.49; 122.02; 123.19; 129.59; 130.20; 130.43; 144.17; 144.94; 150.40;155.92; 158.31; 159.26; 172.07.

IR (KBr): 1251; 1604; 1655; 1735.

EXAMPLE 22 9,11-dichloro-5-ethyl-4,5-dihydro-5-hydroxy-1H-oxepino[3',4':6,7] indolizino[1,2-b] quinoline-3,15 (4H,13H)-dione

This compound is obtain from 3,5-dichloroaniline according to the methodillustrated by the stages 11.i, 11.j, and 11.k of example 11. Yellowsolid, m.p. >250° C.

NMR-¹ H (DMSO): 0.85 (t, 3H); 1.85 (q, 2H) ; 3.07 (d, 1H); 3.45 (d, 1H);5.30 (s, 2H); 5.41 (d, 1H); 5.55 (d, 1H); 6.08 (s, 1H); 7.41 (s, 1H);8.05(s, 1H); 8.21(s,1H); 8.91(s,1H).

NMR-C¹³ (DMSO): 8.93; 36.45; 42.51; 51.03; 61.39; 73.25; 100.62; 123.55;124.63; 127.60; 128.08; 128.56; 132.06; 132.19; 134.53; 143.77; 148.80;154.88; 155.82; 159.13; 171.98.

IR (KBr): 1064; 1275; 1586; 1651; 1743.

EXAMPLE 23 5-ethyl-9-fluoro-4,5-dihydro-5-hydroxy-10-methyl-1H-oxepino[3',4':6,7] indolizino [1,2-b] quinoline-3,15 (4H,13H)-dione

This compound is obtained from 3-fluoro-4-methylaniline according to themethod illustrated by the stages 11.i, 11.j, and 11.k of example 11.Yellow solid, m.p. >250° C.

NMR-¹ H (DMSO):: 0.89 (t, 3H); 1.85 (q, 2H); 2.49 (s, 3H); 3.08 (d, 1H);3.49 (d, 1H); 5.21 (s, 2H); 5.39 (d, 1H); 5.51 (d, 1H); 6.05 (s, 1H);7.39 (s, 1H); 7.87(d, 1H); 8.05(d,1H); 8.61(s,1H).

NMR-C¹³ (DMSO): 8.40; 15.14; 36.45; 42.52; 50.60; 61.41; 73.28; 99.71;112.00; 122.66; 125.38; 127.66; 129.59; 130.28; 144.49; 147.88; 152.88;155.85; 159.18; 162.25; 172.02.

IR (KBr): 1054; 1580; 1651; 1760.

EXAMPLE 24 5-ethyl-10-fluoro-4,5-dihydro-5-hydroxy-1H-oxepino[3',4':6,7] indolizino [1,2-b] quinoline-3,15 (4H,13H)-dione

This compound is obtained from 4-fluoroaniline according to the methodillustrated by the stages 11.i, 11.j, and 11.k of example 11. Whitesolid, m.p. >250° C.

NMR-¹ H (DMSO): 0.85 (t, 3H); 1.85 (q, 2H); 3.07 (d, 1H); 3.45 (d, 1H);5.29 (s, 2H); 5.39 (d, 1H); 5.55(d, 1H); 6.30 (s, 1H); 7.39 (s, 1H);7.80(q, 1H); 7.99(q,1H); 8.23(q,1H); 8.68 (s,1H).

NMR-C¹³ (DMSO): 8.40; 36.46; 42.48; 50.66; 61.41; 73.31; 99.68; 111.83;122.75; 128.93; 130.93; 131.22; 131.93; 144.46; 145.27; 152.60; 155.89;159.21; 172.04.

IR (KBr): 1209; 1589; 1659; 1739.

EXAMPLE 25 10-chloro-5-ethyl-4,5-dihydro-5-hydroxy-1H-oxepino[3',4':6,7] indolizino [1,2-b] quinoline-3,15 (4H,13H)-dione

This compound is obtained from 4-chloroaniline according to the methodillustrated by the stages 11.i, 11.j, and 11.k of example 11. Yellowsolid, m.p. >250° C.

NMR-¹ H (DMSO): 0.85 (t, 3H); 1.85 (q, 2H) ; 3.07 (d, 1H); 3.47 (d, 1H);5.25 (s, 2H); 5.39 (d, 1H); 5.51(d, 1H); 6.05(s, 1H); 7.39 (s, 1H);7.89(d, 1H); 8.19(d,1H); 8.29(s,1H); 8.67 (s,1H).

NMR-C¹³ (DMSO): 8.40; 36.46; 42.48; 50.70; 61.42; 73.31; 100.00; 122.96;127.31; 127.42; 128.87; 131.11; 132.12; 144.34; 146.53; 153.38; 155.88;159.20; 172.04.

IR (KBr): 1069; 1483; 1606; 1741.

EXAMPLE 26 10-chloro-5-ethyl-9-fluoro-4,5-dihydro-5-hydroxy-1H-oxepino[3',4':6,7] indolizino [1,2-b] quinoline-3,15 (4H,13H)-dione

This compound is obtained from 4-chloro-3-fluoroaniline according to themethod illustrated by the stages 11.i, 11.j, and 11.k of example 11.Yellow solid, m.p. >250° C.

NMR-¹ H (DMSO): 0.85 (t, 3H); 1.85 (q, 2H); 3.07 (d, 1H); 3.45 (d, 1H);5.25 (s, 2H); 5.39 (d, 1H); 5.51(d, 1H); 6.05(s, 1H); 7.40 (s, 1H);8.20(d, 1H); 8.40(d,1H); 8.68 (s,1H).

NMR-C¹³ (DMSO): 8.38; 36.47; 42.58; 50.71; 61.40; 73.26; 99.99; 133.59;123.09; 124.28; 127.74; 130.64; 131.31; 144.13; 145.08; 153.57; 154.13;155.84; 156.61; 159.14; 172.00.

IR (KBr): 1488; 1583; 1655; 1743.

EXAMPLE 27 5,12-diethyl-4,5-dihydro-5,10-dihydroxy-11-morpholinomethyl-1H-oxepino [3',4':6,7] indolizino [1,2-b] quinoline-3,15(4H,13H)-dione

This compound is obtained from morpholine according to the methodillustrated in the example 15a. White solid, m.p. >250° C.

NMR-¹ H (DMSO): 0.85 (t, 3H); 1.87 (q, 2H); 2.53 (s, 4H); 3.03 (d, 1H);3.45 (d, 1H); 3.57 (s,4H); 4.02 (s, 2H); 5.01 (s, 2H); 5.38 (d, 1H);5.52(d, 1H); 6.0(se, 1H); 7.30 (s, 1H); 7.42(d, 1H); 7.95 (d,1H);8.82(s,1H).

NMR-C¹³ (DMSO): 8.45; 3.49; 42.58; 53.04; 61.44; 66.33; 73.33; 98.81;113.78; 121.81; 122.74; 126.80; 129.05; 129.91; 143.72; 145.07; 149.24;155.06; 156.92; 159.28; 172.08.

IR (KBr): 1515; 1595; 1654; 1736.

EXAMPLE 285,12-diethyl-9-fluoro-4,5-dihydro-5-hydroxy-10-methoxy-1H-oxepino[3',4':6,7] indolizino [1,2-b] quinoline-3,15 (4H,13H)-dione 28.a.5-fluoro-4-methoxy-2-propionylaniline

(This product is obtained according Sugasawa T., Toyoda T., Adachi M.,Saskura K., J. Am. Chem. Soc., 100 (1978), pp. 4842-4852). A solution of3-fluoro-4-methoxyaniline (20 g, 142 mmol) in anhydrous dichloromethane(200 ml), under an argon atmosphere and at 0° C., is added drop-wise toboron trichloride (1 M in heptane, 156 ml, 156 mmol). The pinksuspension thus obtained is maintained under agitation 5 min. Thenpropionitril (33 ml, 420 mmol) followed by aluminum trichloride (20.9 g,156 mmol) is added drop-wise in small portions. The reaction medium isheated to reflux for 3 h, chilled at 0° C., hydrolyzed by adding,carefully, 2N hydrochloric acid (100 ml), then brought to reflux for 45min. After chilling to 0° C., a precipitate is obtained which is thenfiltered, washed with dichloromethane, then recovered in water (300 ml).The aqueous phase is basified to an alkaline pH, extracted withdichloromethane then ethyl acetate. The organic phase is dried (MgSO₄)then evaporated resulting in a crude product which is purified by columnchromatography (SiO₂, AcOEt/Hpt: 1/99 to 20/80). 15.3 g of a yellowsolid is obtained.

NMR-1H (CDCl3): 1.20 (t, 3H); 2.92 (q, 2H); 3.83 (s, 3H); 6.2 (s, 2H);6.40 (d, 2H); 7.32 (d, 2H).

IR (KBr): 857; 1148; 1240; 1561; 1583; 1662.

28.b. ethyl 4-ethyl-7-fluoro-2-hydroxy-6-methoxy-3-quinolinecarboxylate

A solution of 5-fluoro-4-methoxy-2-propionylaniline (15.3 g, 77.5 mmol)and triethylamine (13.9 ml, 100 mmol) in anhydrous acetonitrile (110ml), under argon and at 0° C., is added drop-wise to a solution ofethylmalonyl chloride (12.9 ml, 100 mmol) in anhydrous acetonitrile (30ml). The reaction medium is left to return to ambient temperature, asolution of sodium ethylate (obtained with 1.8 g, 78 mmol, of sodium in80 ml of ethanol), then is left to agitate for 12 hours at ambienttemperature. The reaction mixture is poured into ice water (100 ml) andagitated for 2 hours, then the precipitate is filtered and washed withwater, ethanol and ether. 19.4 g of a white solid is obtained.

NMR-¹ H (CDCl₃): 1.25 (m, 6H); 2.78 (q, 2H); 3.92 (s, 3H); 4.30 (q, 2H);7.15 (d,2H); 7.40 (d, 2H); 11.93 (s, 1H).

IR (KBr): 786; 1083; 1410; 1521; 1644; 1725.

28.c. ethyl 2-chloro-4-ethyl-7-fluoro-6-methoxy-3-quinolinecarboxylate

A suspension of ethyl4-ethyl-7-fluoro-2-hydroxy-6-methoxy-3-quinolinecarboxylate (19.4 g,0.066 mol) in phosphoryl chloride (243 ml) is brought to reflux for 6 h.The phosphoryl chloride is distilled. The reaction mixture is pouredinto ice water. Recover with dichloromethane to make soluble. Theorganic phase is washed with water, then with a saturated solution ofsodium chloride. The organic phase is dried on magnesium sulfate and thesolvent is evaporated. The residue is suspended in ether and filter thenon-converted starting product (4 g) is filtered. The filtrate isevaporated and the residue is purified by column chromatography (SiO2,AcOEt/Hpt: 5/95 to 20/80). 10.9 g of a white solid is obtained.

NMR-1H (CDCl3): 1.30 (t, 3H); 1.39 (t, 3H); 3.08 (q, 2H); 4.09 (s, 3H);4.49 (q, 2H); 7.64 (d, 2H); 7.86 (d, 2H).

IR (KBr): 865; 1016; 1082; 1190; 1224; 1253; 1272; 1508; 1571; 1732.

28.d. 2-chloro-4-ethyl-7-fluoro-6-methoxy-3-quinolinemethanol

A solution of ethyl2-chloro-4-ethyl-7-fluoro-6-methoxy-3-quinolinecarboxylate (10.8 g, 35mmol) in anhydrous dichloromethane (200 ml) is treated drop-wise atambient temperature under inert atmosphere with diisobutylaluminumhydride (1M in dichloromethane, 65 ml, 65 mmol), then heated to 40° C.for 4 h. Chilled to 0° C., a 20% aqueous solution of Rochelle salt isadded carefully and dichloromethane (200 ml) and the agitation ismaintained for 1 h. Decant, wash three times with water, and dry theorganic phase on magnesium sulfate and evaporate the solvent. Theresidue is purified by column chromatography (SiO2, AcOEt/Hpt: 5/95 to50/50). 6 g of a white solid is obtained.

NMR-¹ H (CDCl3): 1.28 (t, 3H); 3.25 (q, 2H); 4.04 (s, 3H); 4.77 (d, 2H);5.27 (t, 1H); 7.55 (d, 2H); 7.73 (d, 2H).

IR (KBr): 840; 864; 1023; 1232; 1267; 1317; 1444; 1511; 1569.

28.e. 5,12-diethyl-9-fluoro-4,5-dihydro-5-hydroxy-10-methoxy-1H-oxepino[3',4':6,7] indolizino [1,2-b] quinoline-3,15 (4H,13H)-dione

2-chloro-4-ethyl-7-fluoro-6-methoxy-3-quinolinemethanol is coupled tocompound (M) as described in stage 11.j. of example 11. The resultantcoupling product is cyclized according to the procedure of stage 11.k. Ayellow solid is obtained, m.p. >275° C.

NMR-¹ H (CF₃ COOD): 1.07 (m, 3H); 1.62 (m, 3H); 2.27 (m, 2H); 3.44(d,1H); 3.54 (m, 2H); 3.91 (d, 1H); 4.25 (s, 3H); 5.60 (d, 1H); 5.74 (s,2H); 5.98 (d, 1H); 7.85 (m, 1H); 8.16 (m, 1H); 8.31 (s, 1H).

NMR-C¹³ (CF3COOD): 9.03; 14.20; 26.68; 38.77; 43.98; 53.79; 58.27;64.73; 77.93; 106.85; 109.24; 110.15; 128.99; 129.20; 131.61; 137.32;141.23; 144.13; 154.79; 158.32; 160.25; 160.81; 179.30.

IR (KBr): 1013; 1068; 1265; 1466; 1514; 1601; 1655; 1748.

EXAMPLE 29 5-ethyl-4,5-dihydro-5-hydroxy-12-methyl-1H-oxepino[3',4':6,7] indolizino [1,2-b] quinoline-3,15 (4H,13H)-dione

2-acetylaniline is applied to the procedures described by the examples28.b., 28.c. and 28.d. resulting in2-chloro-4-methyl-3-quinolinemethanol. The latter is coupled to compound(M) as describe in stage 11.j. of example 11. The resultant couplingproduct is cyclized according to the procedure of stage 11.k. A yellowsolid is obtained, m.p. >260° C.

NMR-¹ H (DMSO): 0.87 (t, 3H); 1.87 (q, 2H); 2.78 (s, 3H); 2.80 (d, 1H);3.55 (d, 1H); 5.27 (s, 2H); 5.42 (d, 1H); 5.52 (d, 1H); 6.04 (s, 1H);7.39 (s, 1H); 7.75 (t, 1H); 7.88 (t, 1H); 8.13 (d, 1H); 8.25 (d, 1H).NMR-C¹³ (DMSO): 8.23; 36.26; 42.36; 62.00; 73.11; 78.65; 79.13; 79.25;99.52; 122.36; 124.30; 127.67; 129.54; 129.55; 129.56; 140.11; 145.06;148.07; 152.00; 155.79; 159.09; 171.89.

IR (KBr): 1649; 1751; 3404.

EXAMPLE 309-chloro-5-ethyl-4,5-dihydro-5-hydroxy-10-methoxy-12-(4-methylpiperazinomethyl)-1H-oxepino [3',4':6,7] indolizino [1,2-b]quinoline-3,15 (4H,13H)-dione 30.a.5-chloro-2-chloroacetyl-4-methoxyaniline

This product is obtained according to Sugasawa T., Toyoda T., Adachi M.,Sasakura K., J. Am. Chein. Soc., 100 (1978), p. 4842-4852). A molarsolution of boron trichloride in hexane (164 ml, 164 mmol),chloroacetonitrile (11.4 ml, 180 mmol), and a molar solution ofdiethylaluminum chloride in hexane (164 ml, 164 mmol) is added drop-wisein succession to a solution of 3-chloro-4-methoxy-aniline (23.6 g, 150mmol) in an inert atmosphere at 0° C. The reaction medium is heated toreflux for 1 h then cooled to 0° C., hydrolized by adding carefully 2Nhydrochloric acid (90 ml), then maintained at reflux for 1 h. It ischilled again and a concentrated solution of sodium bicarbonate is addedto pH 14. The solution is extracted with ethyl acetate, the organicphase is washed in water, then in salt water. Dry it on magnesuimsulfate, filter and evaporate under reduced pressure. The residue isrecovered in isopentane, decanted, then recover what is insoluble in theminimum amount of isopropyl ether, and isopentane is added toprecipitate the product. Filter and dry under vacuum. 17.26 g of a brownsolid are obtained.

NMR-¹ H (CDCl₃): 3.82 (s, 3H); 4.60 (s, 2H); 6.11 (s, 2H); 6.78 (s, 1H);7.11 (s, 1H).

30.b. ethyl7-chloro-4-chloromethyl-2-hydroxy-6-methoxy-3-quinolinecarboxylate

Ethylmalonyl chloride (17 ml, 131 mmol) is added drop-wise to a solutionof 5-chloro-2-chloroacetyl-4-methoxyaniline (17 g, 73 mmol) andtriethylamine (18.5 ml, 131 mmol) in anhydrous acetonitrile (310 ml),under argon and at 0° C. Agitate for 2 h at ambient temperature, thenadd drop-wise at 0° C. a solution of sodium ethanolate in ethanol(obtained by 1.88 g, 80 mmol, of sodium in 90 ml of ethanol). Agitatedfor 12 hours at ambient temperature. Add 300 ml of water, agitate foranother 20 minutes. Filter the precipitate; wash with water, ethanol,and ethyl ether. After drying in vacuum, 16.7 g of a yellowish solid isobtained.

NMR-¹ H (DMSO): 1.31(t, 3H); 3.95 (s, 3H); 4.36 (q, 2H); 4.95 (s, 2H);7.46 (s, 1H); 7.49 (s, 1H).

30.c. ethyl 2,7-chloro-4-chloromethyl-6-methoxy-3-quinoline-carboxylate

A suspension of ethyl7-chloro-4-chloromethyl-2-hydroxy-6-methoxy-3-quinolinecarboxylate(116.7 g, 50 mmol) in phosphoryl chloride (100 ml) is brought to refluxfor 6 hours. The phosphoryl chloride is distilled. The residue isrecovered in water and agitated for 30 min. The precipitate is filteredand washed with water until neutralization. The precipitate is recoveredin dichloromethane and with a saturated solution of sodium chloride.Filter on a bed of celite and decant the filtrate. The organic phase iswashed again by a saturated solution of sodium chloride. Dry onmagnesium sulfate, and filter and evaporate under reduced pressure.15.88 g of a brown oil is obtained.

NMR-¹ H (CDCl₃): 1.47 (t, 3H); 4.08 (t, 3H); 4.55 (q, 2H); 4.87 (s, 2H);7.35 (s, 1H); 8.09 (s, 1H).

30.d. ethyl2,7-chloro-6-methoxy-4-(4-methylpiperazinomethyl)-3-quinolinecarboxylate

A mixture of ethyl2,7-chloro-4-chloromethyl-6-methoxy-3-quinoline-carboxylate (6.9 g, 20mmol) and N-methylpiperazine (9 ml, 80 mmol) is heated to 60° C. for 30min. The reaction mass is diluted in water and extracted with ethylacetate. Decant and wash the organic phase with water. Dry on magnesiumsulfate, filter and evaporate under reduced pressure. Recover theresidue in water, agitate 15 minutes, filter, wash with water and dry ina vacuum. Purify the residue by column chromatography (SiO₂, MeOH/CH₂Cl₂ : 5/95 to 8/92). 6.7 g of a beige solid product is obtained.

NMR-¹ H (CDCl₃): 1.45 (t, 3H); 2.28 (s, 3H); 2.35-2.70(m, 8H); 3.86 (s,2H); 4.04 (s, 3H); 4.48 (q, 2H); 7.77 (s, 1H); 8.05 (s, 1H).

30.e.2,7-chloro-6methoxy-4-(4-methylpiperazinomethyl)-3-quinolinemethanol

Ethyl2,7-chloro-6-methoxy-4-(4-methylpiperazinomethyl)-3-quinoline-carboxylate(6 g, 14.5 mmol) is dissolved in methylene chloride (120 ml). A molarsolution of diisobutyl aluminum hydride in methylene chloride (60 ml, 60mmol) is added slowly. Agitate one hour at ambient temperature. Pour thereaction mass slowly into 300 ml of a 20% Rochelle salt solution.Agitate one hour; filter on celite; decant, wash the organic phase witha saturated solution of sodium chloride. Dry on magnesium sulfate,filter and evaporate under reduce pressure. Recover the solid withisopropyl ether, filter and dry in vacuum. 4.3 g of the sought compoundis obtained in the form of a yellow solid.

NMR-¹ H (CDCl₃): 2.27 (s, 3H); 2.30 - 2.80(m, 8H); 4.03 (s, 3H); 4.08(s, 2H); 4.96 (s, 2H); 5.95 (s, 1H); 7.37 (s, 1H); 8.05 (s, 1H).

30.f. 9-chloro-5-ethyl-4,5-dihydro-5-hydroxy-10-methoxy-12-(4-methylpiperazinomethyl)-1H-oxepino [3',4':6,7] indolizino [1,2-b]quinoline-3,15 (4H,13H)-dione

2,7-chloro-6methoxy-4-(4-methylpiperazinomethyl)-3-quinoline-methanol iscoupled to compound (M) as described in stages 11.j. of example 11. Theresultant coupling product is cyclized according to the procedureoutlined in 11.k. A yellow solid is obtained, m.p. >250° C.

NMR-¹ H (DMSO): 0.87 (t, 3H); 1.84 (q, 2H); 2.53 (s, 4H); 3.08 (d, 1H);3.47 (d, 1H); 3.58 (s, 4H); 4.06 (s, 5H); 5.30 (s, 2H); 5.42 (q, 2H);6.03 (s, 1H); 7.31 (s, 1H); 7.91 (s, 1H); 8.16 (s, 1H).

NMR-C¹³ (DMSO): 8.42; 36.53; 50.65; 53.30; 56.67; 62.00; 66.50; 73.32;99.31; 104.86; 122.32; 126.94; 126.70; 129.83; 130.44; 138.89; 144.22;144.85; 151.05; 153.17; 155.92; 159.19; 172.06.

IR (KBr): 862; 1063; 1116; 1248; 1595; 1655; 1744; 3449.

EXAMPLE 31 9-chloro-5-ethyl-4,5-dihydro-5-hydroxy-10-methoxy-12morpholinomethyl-1H-oxepino [3',4':6,7] indolizino [1,2-b]quinoline-3,15 (4H,13H)-dione

Apply 3-chloro-4-methoxyaniline to the procedure described in theexamples 30.a., 30.b. and 30.c. resulting in ethyl2,7-dichloro-4-chloromethyl-6-methoxy-3-quinolinecarboxylate which istreated according to the procedure in example 30.d., using morpholine inplace of N-methylpiperazine, then reduce the solution according to themethod in example 30.e. in corresponding quinolinemethanol. The latteris coupled to compound (M) as described in step 11.j. of example 11. Theresultant coupling product is cyclized according to the procedureoutlined in 11.k. A beige solid is obtained, m.p. >250° C.

NMR-¹ H (DMSO): 0.87 (t, 3H); 1.84 (q, 2H); 2.15 (s, 3H); 2.32 (s, 4H);2.50 (s, 4H); 3.08 (d, 1H); 3.47 (d, 1H); 4.06 (s, 5H); 5.29 (s, 2H);5.46 (q, 2H); 6.06 (s, 1H); 7.31 (s, 1H); 7.92 (s, 1H); 8.17 (s, 1H).

NMR-C¹³ (DMSO): 8.42; 36.51; 42.57; 45.93; 50.66; 52.83; 55.05; 56.09;56.72; 61.44; 73.29; 99.30; 104.89; 122.32; 126.89; 127.63; 129.85;130.16; 138.78; 144.18; 144.81; 151.03; 153.01; 155.10; 159.17; 172.07.

IR (KBr): 1055; 1252; 1596; 1655; 1747; 3449.

EXAMPLE 32 5-ethyl-4,5-dihydro-5-hydroxy-12-(4-methylpiperazinomethyl)-1H-oxepino [3',4':6,7] indolizino [1,2-b]quinoline-3,15 (4H,13H)-dione

Apply aniline to the procedures described in the examples 30.a., 30.b.and 30.c. resulting in ethyl2-chloro-4-chloromethyl-3-quinolinecarboxylate which is treatedaccording to the procedure in example 30.d. with N-methylpiperazine,then reduced according to the method in example 30.e. in correspondingquinolinemethanol. The latter is coupled to compound (M) as described instep 11.j. of example 11. The resultant coupling product is cyclizedaccording to the procedure outlined in 11.k. A yellow solid is obtained,m.p. >260° C.

NMR-¹ H (DMSO): 0.86 (t, 3H); 1.87 (q, 2H); 2.14 (s, 3H); 2.32-2.60 (m,8H); 3.05 (d, 1H); 3.48 (d, 1H); 4.09 (q, 2H); 5.42 (d, 1H), 5.52 (d,1H); 6.03 (se, 1H); 7.40 (s, 1H); 7.72 (t, 1H); 7.85 (t, 1H); 8.16 (d,1H); 8.45 (d, 1H).

IR (KBr): 1652; 1735; 3424.

EXAMPLE 33 5-ethyl-4,5-dihydro-5-hydroxy-12-piperidinomethyl-1H-oxepino[3',4':6,7] indolizino [1,2-b] quinoline-3,15 (4H,13H)-dione

Apply aniline to the procedures described in the examples 30.a., 30.b.and 30.c. resulting in ethyl2-chloro-4-chloromethyl-3-quinolinecarboxylate which is treatedaccording to the procedure in example 30.d. by using piperidine in placeof N-methylpiperazine, then reduced according to the method in example30.e. in corresponding quinolinemethanol. The latter is coupled tocompound (M) as described in step 11.j. of example 11. The resultantcoupling product is cyclized according to the procedure outlined in11.k. A yellow solid is obtained, m.p. >260° C.

NMR-¹ H (DMSO): 0.86 (t, 3H); 1.40 (se, 2H); 1.48 (se, 4H); 1.87 (q,2H); 2.50 (s,4H); 3.05 (d, 1H); 3.48 (d, 1H); 4.04 (q, 2H); 5.33 (s,2H); 5.42 (d, 1H); 5.51 (d, 1H); 6.07 (se, 1H); 7.75 (t, 1H); 7.85(t,1H); 8.15 (d, 1H); 8.45 (d, 1H).

NMR-C¹³ (DMSO): 8.47; 23.50; 25.82; 36.50; 42.50; 50.68; 54.47; 58.00;61.42; 73.35; 99.55; 122.61; 125.31; 127.58; 129.54; 129.55; 129.56;129.57; 140.49; 144.95; 148.63; 152.41; 155.90; 159.23; 172.07.

IR (KBr): 1659; 1727; 3408.

EXAMPLE 34 5-ethyl-4,5-dihydro-5-hydroxy-12-morpholinomethyl-1H-oxepino[3',4':6,7] indolizino [1,2-b] quinoline-3,15 (4H,13H)-dione

Apply aniline to the procedures described in the examples 30.a., 30.b.and 30.c. resulting in ethyl2-chloro-4-chloromethyl-3-quinolinecarboxylate which is treatedaccording to the procedure in example 30.d. by using morpholine in placeof N-methylpiperazine, then reduced according to the method in example30.e. in corresponding quinolinemethanol. The latter is coupled tocompound (M) as described in step 11.j. of example 11. The resultantcoupling product is cyclized according to the procedure outlined in11.k. A yellow solid is obtained, m.p. >260° C.

NMR-¹ H (DMSO): 0.86 (t,3H); 1.87 (q, 2H); 3.05 (d, 1H); 3.30 (s, 4H);3.49 (d, 1H); 3.55 (se, 4H); 4.10 (q, 2H); 5.35 (s, 2H); 5.40 (d, 1H);5.54 (d.1H); 6.04 (s, 1H); 7.72 (t, 1H); 7.85 (t, 1H); 8.16 (d, 1H);8.47 (d, 1H).

NMR-C¹³ (DMSO): 8.42; 36.51; 42.57; 50.68; 53.51; 56.06; 61.42; 66.41;73.34; 99.56; 122.64; 125.25; 127.56; 129.81; 139.55; 144.92; 148.62;152.39; 155.89; 159.21; 172.05.

IR (KBr): 1657; 1729; 3347.

EXAMPLE 355-etbyl-10-fluoro-4,5-dihydro-5-hydroxy-12-(4methylpiperazinomethyl)-1H-oxepino[3',4':6,7] indolizino [1,2-b] quinoline-3,15 (4H,13H)-dione

Apply 4-fluoroaniline to the procedures described in the examples 30.a.,30.b. and 30.c. resulting in ethyl2-chloro-4-chloromethyl-6-fluoro-3-quinolinecarboxylate which is treatedaccording to the procedure in example 30.d. with N-methylpiperazine,then reduced according to the method in example 30.e. in correspondingquinolinemethanol. The latter is coupled to compound (M) as described instep 11.j. of example 11. The resultant coupling product is cyclizedaccording to the procedure outlined in 11.k. A yellow solid is obtained,m.p. >275° C.

NMR-¹ H (DMSO): 0.87 (t, 3H); 1.85 (q, 2H); 2.15 (s, 3H); 2.31 (m, 4H);2.50 (m, 4H); 3.07 (d, 1H); 3.48 (d, 1H); 4.04 (m, 2H); 5.31 (s, 2H);5.40 (d, 1H); 5.53 (d, 1H); 6.05 (s, 1H); 7.38 (s, 1H); 7.77 (m, 1H);8.19 (m, 2H).

NMR-C¹³ (DMSO): 8.43; 36.51; 42.54; 45.89; 50.67; 52.92; 54.93; 55.92;73.32; 99.56; 122.69; 130.43; 132.40; 139.69; 144.70; 145.84; 152.19;155.90; 159.17; 172.05.

IR (KBr): 836; 1051; 1217; 1291; 1612; 1662; 1726.

EXAMPLE 365-ethyl-10-fluoro-4,5-dihydro-5-hydroxy-12morpholinomethyl-1H-oxepino[3',4':6,7] indolizino [1,2-b] quinoline-3,15 (4H,13H)-dione

Apply 4-fluoroaniline to the procedures described in the examples 30.a.,30.b. and 30.c. resulting in ethyl2-chloro-4-chloromethyl-6-fluoro-3-quinolinecarboxylate which is treatedaccording to the procedure in example 30.d. by using morpholine in placeof N-methylpiperazine, then reduce according to the method in example30.e. in corresponding quinolinemethanol. The latter is coupled tocompound (M) as described in step 11.j. of example 11. The resultantcoupling product is cyclized according to the procedure outlined in11.k. A beige solid is obtained, m.p. >250° C.

NMR-¹ H (DMSO): 0.87 (m, 3H); 1.85 (m, 2H); 2.51 (m,4H); 3.06 (d, 1H);3.48 (d, 1H); 3.56 (m, 4H); 4.05 (m, 2H); 5.34 (s, 2H); 5.40 (d, 1H);5.53 (d, 1H); 6.04 (s, 1H); 7.38 (s, 1H); 7.77 (m, 1H); 8.21 (m, 2H).

NMR-C¹³ (DMSO): 8.40; 36.47; 42.52; 50.59; 53.40; 56.14; 61.44; 66.41;73.29; 99.58; 109.05; 109.28; 120.11; 120.37; 122.68; 128.53; 130.53;132.43; 139.13; 144.62; 145.79; 152.07; 155.94; 159.14; 161.59; 172.04.

IR (KBr): 834; 860; 1061; 1118; 1215; 1286; 1516; 1609; 1658; 1734.

EXAMPLE 375-ethyl-9-fluoro-4,5-dihydro-5-hydroxy-10-methyl12-(4-methylpiperazinomethyl)-1H-oxepino[3',4':6,7] indolizino [1,2-b] quinoline-3,15 (4H,13H)-dione

Apply 3-fluoro-4-methylaniline to the procedures described in theexamples 30.a., 30.b. and 30.c. resulting in ethyl2-chloro-4-chloromethyl-7-fluoro-6-methyl-3quinolinecarboxylate which istreated according to the procedure in example 30.d. withN-methylpiperazine, then reduced according to the method in example30.e. in corresponding quinolinemethanol. The latter is coupled tocompound (M) as described in step 11.j. of example 11. The resultantcoupling product is cyclized according to the procedure outlined in11.k. A yellow solid is obtained, m.p. >260° C.

NMR-¹ H (CDCl₃): 1.00 (t, 3H); 2.00 (q, 2H); 2.35 (s, 3H); 2.50 (s, 3H);2.61 (m, 8H); 3.33 (d, 1H); 3.39 (d, 1H); 3.97 (d, 1H); 4.07 (d, 1H);5.17 (d, 1H); 5.38 (d, 1H); 5.52 (d, 1H); 5.63 (d, 1H); 7.13 (d, 1H);7.28 (s, 1H); 7.99 (d, 1H).

IR (KBr): 1652; 1747; 3430.

EXAMPLE 385-ethyl-9-fluoro-4,5-dihydro-5-hydroxy-10-methyl-12morpholinomethyl-1H-oxepino[3',4':6,7] indolizino [1,2-b] quinoline-3,15 (4H,13H)-dione

Apply 3-fluoro-4-methylaniline to the procedures described in theexamples 30.a., 30.b. and 30.c. resulting in ethyl2-chloro-4-chloromethyl-7-fluoro-6-methyl-3quinolinecarboxylate which istreated according to the procedure in example 30.d. by using morpholinein place of N-methylpiperazine, then reduce according to the method inexample 30.e. in corresponding quinolinemethanol. The latter is coupledto compound (M) as described in step 11.j. of example 11. The resultantcoupling product is cyclized according to the procedure outlined in11.k. A yellow solid is obtained, m.p. >260° C.

NMR-¹ H (DMSO+CDCl₃): 1.00 (t, 3H); 2.02 (q, 2H); 2.57 (s, 3H); 2.60 (s,4H); 3.23 (d, 1H); 3.45 (d, 1H); 3.75 (s, 4H); 4.11 (s, 2H); 5.44 (s,2H); 5.47 (d, 1H); 5.65 (d, 1H); 7.62 (s, 1H); 7.73 (d, 1H); 8.24 (d,1H).

NMR-C¹³ (CF₃ CO₂ D): 8.35; 13.93; 16.01; 22.24; 25.29; 38.18; 43.42;54.19; 56.04; 56.74; 64.16; 65.09; 77.48; 108.29; 108.57; 128.07;128.70; 129.90; 135.64; 138.03; 139.86; 141.10; 141.56; 147.78; 158.30;161.87; 178.72.

IR (KBr): 117; 1609; 1654; 1750; 3437.

EXAMPLE 395-ethyl-9-fluoro-4,5-dihydro-5-hydroxy-10-methyl-12piperidinomethyl-1H-oxepino[3',4':6,7] indolizino [1,2-b] quinoline-3,15 (4H,13H)-dione

Apply 3-fluoro-4-methylaniline to the procedures described in theexamples 30.a., 30.b. and 30.c. resulting in ethyl2-chloro-4-chloromethyl-7-fluoro-6-methyl-3-quinolinecarboxylate whichis treated according to the procedure in example 30.d. by usingpiperidine in place of N-methylpiperazine, then reduce according to themethod in example 30.e. in corresponding quinolinemethanol. The latteris coupled to compound (M) as described in step 11.j. of example 11. Theresultant coupling product is cyclized according to the procedureoutlined in 11.k. A yellow solid is obtained, m.p. >260° C.

NMR-¹ H (CF₃ CO₂ D): 1.09 (s, 3H); 1.70 (t, 1H); 2.03 (m, 5H); 2.25 (s,2H); 2.70 (s,3H); 3.54 (d, 3H); 3.88 (d, 1H); 4.01 (se,2H); 5.30 (q,2H); 5.65 (d, 1H); 5.96 (d, 1H); 6.10 (s, 2H); 8.16 (d, 1H); 8.35 (s,1H); 8.61 (s, 1H).

NMR-C¹³ (CF₃ CO₂ D): 8.47; 16.07; 20.93; 22.18; 24.76; 38.28; 43.53;54.30; 56.12; 58.33; 64.24; 77.56; 108.37; 111.30; 128.20; 129.02;129.98; 135.60; 138.29; 139.90; 141.60; 142.26; 147.57; 158.28; 161.90;167.63; 170.31; 178.82.

IR (KBr) :1605; 1657; 1728; 3399.

EXAMPLE 40 8-ethyl-2,3,8,9-tetrahydro-8-hydroxy-16-(4-methylpiperazinomethyl)-10H,12H-(1,4) dioxino (2,3-g) oxepino [3',4':6,7]indolizino [1,2-b] quinoline-10,13[15H]-dione

Apply 3,4-ethylenedioxyaniline to the procedures described in theexamples 30.a., 30.b. and 30.c. resulting in ethyl2-chloro-4-chloromethyl-6,7-ethylenedioxy-3quinolinecarboxylate which istreated according to the procedure in example 30.d. withN-methylpiperazine, then reduced according to the method in example30.e. in corresponding quinolinemethanol. The latter is coupled tocompound (M) as described in step 11.j. of example 11. The resultantcoupling product is cyclized according to the procedure outlined in11.k. A yellow solid is obtained, m.p. >260° C.

NMR-¹ H (DMSO): 0.92 (t, 3H); 1.89 (q, 2H); 2.16 (s, 3H); 2.50 (m, 8H);3.12 (d, 1H); 3.50 (d, 1H); 3.95 (s, 2H); 4.47 (s, 4H); 5.19 (q, 2H);5.43 (d, 1H); 5.56 (d, 1H); 7.35 (s, 1H); 7.54 (s, 1H); 7.76 (s, 1H).

NMR-C¹³ (DMSO): 8.45; 24.80; 36.51; 42.48; 45.90; 50.45; 52.98; 54.91;56.10; 61.44; 64.43; 73.30; 99.03; 109.46; 1 13.5 1; 121.95; 123.51;127.76; 137.99; 145.00; 145.14; 145.27; 147.24; 150.53; 155.90; 159.18;172.27; 177.00.

IR (KBr): 1656; 1743; 3422.

EXAMPLE 419-chloro-5-ethyl-10-fluoro-4,5-dihydro-5-hydroxy-12morpholinomethyl-1H-oxepino[3',4':6,7] indolizino [1,2-b] quinoline-3,15 (4H,13H)-dione

Apply 3-chloro-4-fluoroaniline to the procedures described in theexamples 30.a., 30.b. and 30.c. resulting in ethyl2,7-dichloro-4-chloromethyl-6-fluoro-6-3-quinolinecarboxylate which istreated according to the procedure in example 30.d. using morpholine inplace of N-methylpiperazine, then reduced according to the method inexample 30.e. in corresponding quinolinemethanol. The latter is coupledto compound (M) as described in step 11.j. of example 11. The resultantcoupling product is cyclized according to the procedure outlined in11.k. A beige solid is obtained, m.p. >250° C.

NMR-¹ H (CF₃ COOD): 1.09 (t, 3H); 2.30 (m, 2H); 3.50 (d, 1H); 3.90 (d,1H); 3.98 (d, 4H); 4.36 (s, 4H); 5.38 (q, 2H),; 5.64 (d, 1H); 5.96 (d,1H); 6.23 (q, 2H); 8.57 (d, 1H); 8.60 (s, 1H); 8.85 (d, 1H).

NMR-C¹³ (CF₃ COOD): 8.10; 37.80; 43.11; 54.31; 55.78; 63.75; 65.11;77.06; 128.28; 129.55; 130.33; 136.26; 137.11; 138.40; 139.67; 139.85;148.58; 157.54; 159.74; 161.31; 178.00.

IR (KBr) :848; 1042; 1230; 1609; 1658; 1750; 3310; 3387.

EXAMPLE 42 resolution of 5-ethyl-4,5-dihydro-5-hydroxy-1H-oxepino[3',4':6,7]-indolizino [1,2-b] quinoline-3,15 (4H,13H)-dione

A mixture of β-ethyl-β-hydroxy-γ-(8-hydroxymethylindolizino [1,2-b]quinoline-9(11H)-one-7-yl)-propionic acid (19.5 g, 52 mmol) andL-(-)-α-methylbenzylamine (12.12 g, 100 mmol) in absolute ethanol (11)is carried to boiling, hot-filtered and allowed to rest for 68 h. Theprecipitate is filtered and washed with ethanol and ether resulting in9.8 g of a white solid. An analysis by high pressure affinitychromatography on the chiral phase ("HPLC chiral" on a Chiral-AGP column(Chromthech, Stockholm, Sweden) 100×4 mm, 2% acetonitrile eluant in a 10mM, pH 6.9 phosphate buffer, eluant peaks at 4.5 and 7.5 min) revealstwo peaks integrating respectively for 24% and 76% of the total surfaceof the two peaks. Recover the solid in 93% ethanol (350 ml) at reflux,then allow to rest for 48 h. Filter the precipitate then wash withethanol and ether to obtain 4.8 g of a white solid, giving twointegrating peaks respectively for 9% and 91% of the total surface ofthe two peaks by chiral HPLC. Recover the solid in 50% ethanol (48 ml)at reflux then allow to rest for 48 h. Filter the precipitate then washwith ethanol and ether resulting in 2.7 g of a white solid giving twointegrating peaks respectively for 1% and 99% of the total surface ofthe two peaks by chiral HPLC. The resultant salt is treated, enricheddiastereoisomerically, recovered in distilled water (20 ml), with aceticacid (0.35 ml, 6.4 mmol) for 15 min. Filter the precipitate obtained,wash with water, acetone and ether then dry in a vacuum at 80° C.resulting in 1.1 g of a white solid. Recover the latter in absoluteethanol (55 ml) with concentrated hydrochloric acid added (11.5 N, 11ml) resulting in a yellow solution which is agitated at ambienttemperature for 68 h. Filter the precipitate thus obtained and wash withwater, ethanol and ether, then dry in vacuum at 80° C. resulting in 770mg of 5-ethyl-4,5-dihydro-5-hydroxy-1H-oxepino [3',4':6,7]-indolizino[1,2-b] quinoline-3,15 (4H,13H)-dione enantiomerically enriched. Ananalysis by chiral HPLC (Chiral-AGP column, eluted by a 2 to 5% gradientof acetonitrile in a 10 mM at pH 6.9 phosphate buffer, eluant peaks at15 and 20 min) reveal an enantiomeric excess of 98%. Repeat theprocedure described above replacing the L-(-)-α-methylbenzylamine withD-(+)-α-methylbenzylamine. The other enantiomer of5-ethyl-4,5-dihydro-5-hydroxy-1H-oxepino [3',4':6,7]-indolizino[1,2-b]quinoline-3,15 (4H,13H)-dione is thus obtained.

By using the procedures indicated above, the following products may beprepared as well, and are as well part of the invention whichconstitutes the preferred products:

10-benzyloxy-5,12-diethyl-4,5-dihydro-5-hydroxy-1H-oxepino [3',4':6,7]indolizino [1,2-b] quinoline-3,15 (4H,13H)-dione;

5,12-diethyl-4,5-dihydro-5,10,-dihydroxy-1H-oxepino [3',4':6,7]indolizino [1,2-b]quinoline-3,15 (4H,13H)-dione;

5,12-diethyl-4,5-dihydro-5,10-dihydroxy-11-dimethylaminomethyl-1H-oxepino[3',4':6,7] indolizino [1,2-b] quinoline-3,15 (4H,13H)-dione;

5-ethyl-4,5-dihydro-5-hydroxy-9,10-dimethoxy-1H-oxepino [3',4∝:6,7]indolizino [1,2-b] quinoline-3,15 (4H,13H)-dione;

10-bromo-5-ethyl-4,5-dihydro-5-hydroxy-1H-oxepino [3',4':6,7] indolizino[1,2-b] quinoline-3,15 (4H,13H)-dione;

11-bromo-5-ethyl-4,5-dihydro-5-hydroxy-10-methoxy-1H-oxepino [3',4':6,7]indolizino [1,2-b] quinoline-3,15 (4H,13H)-dione;

5-ethyl-12-dimethylaminomethy-4,5-dihydro-5-hydroxy-1H-oxepino[3',4':6,7] indolizino [1,2-b] quinoline-3,15 (4H,13H)-dione;

5-ethyl-4,5-dihydro-5-hydroxy-1H, 3H-cyclopenta [g] oxepino [3',4':6,7]indolizino [1,2-b] quinoline-3,16 (14H)-dione;

7-ethyl-7,8-dihydro-7-hydroxy-16-(4-methylpiperazino methyl)-9H,11H-[1,3] dioxolo [4,5-g] oxepino [3',4':6,7] indolizino [1,2-b]quinoline-9,12 (14H)-dione;

5-ethyl-4,5-dihydro-5-hydroxy-12-(1-imidazylmethyl)-1H-oxepino[3',4':6,7] indolizino [1,2-b ] quinoline-3,15 (4H,13H)-dione;

8-ethyl-8,9-dihydro-8-hydroxy-10H, 12H-oxepino [3',4':6,7] indolizino[1,2-b] pyrido [2,3-g] quinoline-10,13 (15H)-dione;

5-ethyl-4,5,9,10,11,12-hexahydro-5-hydroxy-1H, 3H-benzo [g] oxepino[3',4':6,7] indolizino [1,2-b] quinoline-3,17 (15H)-dione;

8-ethyl-8,9-dihydro-8-hydroxy-14H, 10H, 12H-[1,3] dioxino [4,5-g]oxepino [3',4':6,7] indolizino [1,2-b] quinoline-10,13 [15H]-dione;

7-ethyl-7,8-dihydro-7-hydroxy-1H, 9H, 11H-oxepino [3',4':6,7] indolizino[1,2-b] pyrrolo [2,3-g] quinoline-9,12 (14H)-dione;

7-ethyl-7,8-dihydro-7-hydroxy-1H, 9H, 11H-imidazo [4,5-g] oxepino[3',4':6,7] indolizino [1,2-b] quinoline-9,12 (14H)-dione;

7-ethyl-7,8-dihydro-7-hydroxy-1H, 9H, 11H-oxepino [3',4':6,7] indolizino[1,2-] 1,2,3-triazolo [4,5-g] quinoline-9,12 (14H)-dione;

7-ethyl-7,8-dihydro-7-hydroxy-9H, 11H-oxepino [3',4':6,7] indolizino[1,2-b] thiazolo [4,5-g] quinoline-9,12 (14H)-dione; and

7-ethyl-7,8-dihydro-7-hydroxy-9H, 11H-oxazolo [4,5-g] oxepino[3',4':6,7] indolizino [1,2-b] quinoline-9,12 (14H)-dione.

Pharmacological Study of the Products of the Invention

1. Test of the Activity of the Relaxation Activity of DNA Induced byTopoisomerase 1

All reactions are carried out in a reaction buffer of 20 μl comprised of50 mM of tris-HCl (pH 7.5), 50 mM of KCl, 0.5 mM of dithiothritol, 10 mMof MgCl₂, 0.1 mM of ethyldiamine tetraacetic acid (EDTA), 30 μl/mlbovine albumin serum and 300 ng of coiled pUC19 (Pharmacia Biotech,Orsay, France) with or without compounds to test a definedconcentrations. All of the compounds to be tested are dissolvedinitially in 50 mM of dimethylsulfoxide (DMSO), the other dilutionsbeing made with distilled water. The final concentration in DMSO doesnot exceed 1% (v/v). The reaction is initiated by the addition of oneunit of purified thymus DNA of sheep topoisomerase 1 (Gibco-BRL,Paisley, United Kingdom) and is carried out for 15 minutes at 37° C. Thereactions are stopped by the addition of 3 μl of a mixture comprising 1%dodecyl sodium sulfate, 20 mM EDTA and 500 μg/ml of proteinase K(Boehringer Mannheim, Melan, France). After a supplementary incubationperiod of 30 minutes at 37° C., 2 μl of a charging buffer comprising Na₂HPO₄, 0.3% bromophenol blue and 16% Ficoll are added to samples whichare subjected to 1.2% to 1 V/cm agar gel electrophoresis for 20 hours ina buffer comprising 36 mM pH 7.8 Tris-HCl, 30 mM Na₂ PO₄, 1 mM EDTA and2 μg/ml chloroquin. The gels are dyed with 2 μg/ml ethidium bromide,photographed in 312 nm U.V. light with a photographic appliance and thefluorescent intensity is measured with a bioProfil camera (VilberLourmat, Lyon, France) in order to determine the percentage of relaxedDNA. Each experiment is performed at least three times in duplicate.

In each experiment, coiled DNA plasmid is incubated alone or withtopoisomerase 1. The reaction is completed in the space of 15 minutes,for each compound tested or used as a control the coiled DNA plasmid isincubated in the presence of 500 μM of the compound to be tested with orwithout enzyme plus the compound, at concentrations of 10 μM, 100 μM,200 μM and 500 μM. As indicated in Table 1, examples 2, 3, 4, 9, 10 and11 inhibit the relaxation activity favored by topoisomerase 1 in amanner dependent upon the dose.

                  TABLE 1                                                         ______________________________________                                        Percentage of Relaxed DNA                                                                Concentration μM                                                Example      10     100        200  500                                       ______________________________________                                        Example 2    97.9   78.3       73.2 51.1                                      Example 3    79.9   59.9       55.0 45.7                                      Example 4    99.1   82.2       67.6 32.9                                      Example 9    77.1   33.9       29.7 20.4                                      Example 10   96.9   45.4       26.2 8.7                                       Example 11   65.0   50.3       39.8 31.0                                      ______________________________________                                    

2. Test of Cell Proliferation

Eight tumor cell lines are used in this study L1210 (mouse lymphocyticleukemia), HCT15 and LOVO (adenocarcinoma cell lines of human colon),A549 (carcinoma of human lung), A172, U373, U87 (human glioblastomas).All of these lines are obtained form the American Type CollectionCultures (ATCC), Rockville, Md. The cultures of the L1210 cells insuspension are cultivated in an eagle medium of modified Dulbecco (DMEM)(BioWhitaker, Verviers, Belgium) completed with 10% fetal calf seruminactivated by heating, 2 mM of glutamine, 50 U/ml of penicillin and 50μl/ml of streptomycin. The HT29 cells are cultivated in mono-layercultures in a 5aMcCoy medium (Gibco, Paisley, United Kingdom) completedwith 10% fetal calf serum inactivated by heating then adding 2 mM ofglutamine and 50 μg of gentamycin. The other cells are cultivated in anessential medium of modified Earle (EMEM; Gibco, Paisley, UnitedKingdom) completed with 5% fetal calf serum inactivated by heat, 2 mM ofglutamine, 50 U/ml of penicillin and 50 μg/ml of streptomycin. All ofthe cell lines are cultivated at 37° C. in a humidified atmospherecontaining 95% air and 5% CO₂.

The inhibition of the tumor cell lines is determined by means of an MTTtest, 1500 L1210 cells in a medium of culture (according to the needs ofthe cell medium) are seeded in a well of a microcuvette plaque (level oftissue culture: 96 wells, flat bottom) 24 hours before the treatmentwith the compounds to be tested. For these dose-response studies, thecells are incubated with each of the compounds to be tested or theircorresponding solvent (control) for 48 hours in a final concentrationpool of 1.10⁻¹⁰ to 1.10⁻⁴ M. All the compounds are dissolved just beforeuse in dimethylsulfoxide (DMSO) at a concentration of 50 mM. Otherdilutions of medications are carried out in the culture medium. Thefinal concentration of DMSO never exceeds 0.2% (v/v). As control themedication solutions are replaced by the solvent which is successivelydiluted in the same fashion as the compounds to be tested.

After the incubation period, the marking reagent MTT(3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium; Thiazol bule,Sigma M 565, Sigma, St. Louis Mo.) is added at a final concentration of0.3 mg/ml to each well. The cells are incubated for 4 hours at 37° C. ina humidified atmosphere. This stage permits the mitochondrialdishydrogenase of the live cells to convert into the yellow tetrazoliumsalt MTT in purple formazan crystals. The supernatant is eliminated andthe formed formazan crystals are solubilized with DMSO. The resultantcolored solution is quantified by absorption at 570 nm by using asweeping multi-cuvette spectrophotometer. The data concerning theproliferation is expressed in percentages of live cells in the treatedwells, divided by the live cells in the control. Each point representsthe average of three independent experiments, each experimentrepresenting six determinations.

For the other cell lines (HCT15, LOVO, A549, A172, U373, U87), 1000 to2000 cells are seeded in a well of a plaque of microwells 24 hoursbefore the medicinal treatment. They are incubated with each of thecompounds to be tested or their corresponding solvent (control) for72.hours in a final concentration pool of 1·10⁻¹⁰ to 1·10⁻⁶ M.

The results are expressed in percentages of the proliferationscalculated by the optical density of the cells treated with a medicationdivided by the OD of the control cells (cells treated with DMSO). As isrepresented in Table II, the compounds to be tested inhibited theproliferation of the cells as a function of dose.

                                      TABLE II                                    __________________________________________________________________________    Percentage of Cellular Proliferation                                          Example                                                                             Cell                                                                              Concentration nM                                                    6     lines                                                                             0.1 1   10  100 1000                                                                              10000                                                                              100000                                     __________________________________________________________________________    Example                                                                             L1210                                                                             87.22                                                                             68.92                                                                             42.64                                                                             26.85                                                                             10.83                                                                             2.11 2.20                                       3     HCT15                                                                             86.00                                                                             84.00                                                                             58.00                                                                             44.00                                                                             18.00                                                                             9.00 13.00                                            LOVO                                                                              108.00                                                                            86.00                                                                             54.00                                                                             31.00                                                                             23.00                                                                             10.00                                                                              12.00                                            A549                                                                              132.00                                                                            111.00                                                                            75.00                                                                             39.00                                                                             35.00                                                                             10.00                                                                              11.00                                            A172                                                                              89.00                                                                             101.00                                                                            68.00                                                                             37.00                                                                             27.00                                                                             10.00                                                                              7.00                                             U373                                                                              99.00                                                                             98.00                                                                             40.00                                                                             24.00                                                                             17.00                                                                             13.00                                                                              9.00                                             U87 108.00                                                                            85.00                                                                             42.00                                                                             23.00                                                                             15.00                                                                             5.00 6.00                                       Example                                                                             L1210                                                                             92.14                                                                             97.14                                                                             91.08                                                                             86.28                                                                             46.79                                                                             27.80                                                                              8.09                                       4     HCT15                                                                             91.00                                                                             92.00                                                                             86.00                                                                             78.00                                                                             54.00                                                                             20.00                                                                              7.00                                             LOVO                                                                              80.00                                                                             75.00                                                                             79.00                                                                             69.00                                                                             38.00                                                                             21.00                                                                              5.00                                             A549                                                                              71.00                                                                             76.00                                                                             71.00                                                                             56.00                                                                             26.00                                                                             22.00                                                                              12.00                                            A172                                                                              93.00                                                                             92.00                                                                             98.00                                                                             97.00                                                                             44.00                                                                             31.00                                                                              10.00                                            U373                                                                              86.00                                                                             85.00                                                                             89.00                                                                             63.00                                                                             30.00                                                                             16.00                                                                              2.00                                             U87 98.00                                                                             101.00                                                                            98.00                                                                             74.00                                                                             11.00                                                                             6.00 2.00                                       Example                                                                             L1210                                                                             74.04                                                                             62.05                                                                             44.72                                                                             34.01                                                                             20.20                                                                             4.34 1.58                                       9     HCT15                                                                             94.00                                                                             89.00                                                                             59.00                                                                             35.00                                                                             15.00                                                                             8.00 3.00                                             LOVO                                                                              74.00                                                                             85.00                                                                             44.00                                                                             31.00                                                                             21.00                                                                             4.00 2.00                                             A549                                                                              91.00                                                                             88.00                                                                             50.00                                                                             31.00                                                                             23.00                                                                             5.00 3.00                                             A172                                                                              97.00                                                                             89.00                                                                             44.00                                                                             36.00                                                                             19.00                                                                             3.00 1.00                                             U373                                                                              89.00                                                                             69.00                                                                             24.00                                                                             18.00                                                                             8.00                                                                              3.00 1.00                                             U87 105.00                                                                            72.00                                                                             14.00                                                                             7.00                                                                              4.00                                                                              2.00 6.00                                       Example                                                                             L1210                                                                             91.51                                                                             97.94                                                                             89.28                                                                             67.32                                                                             31.51                                                                             19.78                                                                              3.65                                       10    HCT15                                                                             111.00                                                                            87.00                                                                             103.00                                                                            63.00                                                                             42.00                                                                             17.00                                                                              9.00                                             LOVO                                                                              71.00                                                                             76.00                                                                             77.00                                                                             52.00                                                                             29.00                                                                             18.00                                                                              4.00                                             A549                                                                              71.00                                                                             76.00                                                                             71.00                                                                             56.00                                                                             36.00                                                                             22.00                                                                              7.00                                             A172                                                                              93.00                                                                             92.00                                                                             91.00                                                                             60.00                                                                             39.00                                                                             15.00                                                                              3.00                                             U373                                                                              96.00                                                                             104.00                                                                            87.00                                                                             35.00                                                                             20.00                                                                             10.00                                                                              2.00                                             U87 96.00                                                                             79.00                                                                             89.00                                                                             17.00                                                                             6.00                                                                              5.00 2.00                                       Example                                                                             L1210                                                                             91.99                                                                             81.37                                                                             23.16                                                                             16.38                                                                             5.59                                                                              1.45 1.04                                       11    HCT15                                                                             71.00                                                                             63.00                                                                             45.00                                                                             23.00                                                                             12.00                                                                             9.00 9.00                                             LOVO                                                                              66.00                                                                             42.00                                                                             29.00                                                                             21.00                                                                             8.00                                                                              3.00 3.00                                             A549                                                                              82.00                                                                             44.00                                                                             29.00                                                                             26.00                                                                             4.00                                                                              3.00 2.00                                             A172                                                                              95.00                                                                             53.00                                                                             47.00                                                                             39.00                                                                             12.00                                                                             3.00 2.00                                             U373                                                                              50.00                                                                             30.00                                                                             25.00                                                                             8.00                                                                              2.00                                                                              1.00 2.00                                             U87 40.00                                                                             21.00                                                                             12.00                                                                             6.00                                                                              1.00                                                                              1.00 1.00                                       __________________________________________________________________________

Test of Development in Vivo

The compounds according to the invention are tested in vivo using thelymphoblastic leukemia L1210 cell lines of mice. The tumor cells aremaintained by a series of i.p. means in mice DBA/2 Lafacedu, Lyon,France). In an experimental trial, 10⁶ cells/0.2 ml are injected by i.p.passage into female B6D2F1 mice. The treatment begins day 1 afterinoculation with leukemia and continues to day 4 or to day 8. Thecompounds to be tested are injected i.p. or i.v. in differentconcentrations and the volume injected is adjusted to 0.1 ml/10 g bodyweight. The non-treated mice die between day 9 and day 14, after theinjection of lymphoblastic leukemia L1210 cells and survival of thetreated mice is observed during an extended period of up to 60 days.

The experiment is conducted with 5-ethyl-4,5-dihydro-5-hydroxy-1H-oxepino [3'4':6,7]-indolizine [1,2-b] quinoline-3,15 (4H, 13H)-dione.This compound augments by 50% the duration of life of the mice atconcentrations of between 0.32 and 2.5 mg/kg administered i.p. during 4days and at concentrations of between 0.32 and 5.0 mg/kg administeredi.v. for 8 days.

We claim:
 1. A compound selected from the group consisting ofenaniomeric, racemic mixtures or combinations thereof of a compound ofthe formula ##STR23## wherein R₁ is selected from the group consistingof alkyl of 1 to 6 carbon atoms, haloalkyl of 1 to 6 carbon atoms, loweralkenyl of up to 6 carbon atoms, alkoxyalkyl of 2 to 12 carbon atoms andalkylthio alkyl of 2 to 12 carbon atoms, R₂, R₃ and R₄ are independentlyselected from the group consisting of hydrogen, halogen, haloalkyl of 1to 6 carbon atoms, alkyl of 1 to 6 carbon atoms, alkenyl of 2 to 6carbon atoms, --CN, cyanoalkyl of 1 to 6 carbon atoms, --NO₂, nitroalkylof 1 to 6 carbon atoms, --CONH₂, amidoalkyl of 1 to 6 carbon atoms,hydrazino, hydrazinoalkyl of 1 to 6 carbon atoms, azido, azidoalkyl of 1to 6 carbon atoms, ##STR24## is a substituted or unsubstitutedheterocyclic of 5 to 8 ring atoms with X being ring members selectedfrom the group consisting of --O--, --S--, --CH₂ --, --CH═, --N═,##STR25## --COR₁₀ and unsubstituted or substituted aryl, and aryl loweralkyl wherein the substituent is selected from the group consisting ofalkyl of 1 to 6 carbon atoms, halogen, --NO₂, --NH₂, alkylamino of 1 to6 carbon atoms, haloalkyl of 1 to 6 carbon atoms, hydroxyalkyl of 1 to 6carbon atoms, alkoxy of 1 to 6 carbon atoms and lower alkoxy alkyl of 2to 12 carbon atoms, R₅ is selected from the group consisting ofhydrogen, halogen, halogen lower alkyl, lower alkyl, lower alkoxy, loweralkoxy lower alkyl, lower alkylthio lower alkyl, cycloalkyl, cycloalkyllower alkyl, cyano, cyano alkyl, hydroxy lower alkyl, nitro, (CH₂)_(m)COR₈, (CH₂)_(m) NR₆ COR₈, (CH₂)_(m) NR₆ R₇, (CH₂)_(m) N(CH₃)(CH₂)_(n)NR₆ R₇, (CH₂)_(m) OCOR₈, (CH₂)_(m) OCONR₆ R₇, or substituted orunsubstituted (CH₂)_(m) [N═X], OCO [N═X], (CH₂)_(m) OCO [N═X], aryl oraryl lower alkyl; R₆ and R₇ are independently selected from the groupconsisting of hydrogen, alkyl of 1 to 6 carbon atoms, hydroxyalkyl of 1to 6 carbon atoms, alkylaminoalkyl of 2 to 12 carbon atoms, aminoalkylof 1 to 6 carbon atoms, cycloalkyl of 3 to 7 carbon atoms,cycloalkylalkyl of 4 to 13 carbon atoms, alkenyl of 2 to 6 carbon atoms,alkoxyalkyl of 2 to 12 carbon atoms, haloalkyl of 1 to 6 carbon atomsand substituted and unsubstituted aryl and arylalkyl of 1 to 6 alkylcarbon atoms and the substituent is at least one member of the groupconsisting of halogen, --NO₂, --NH₂, alkyl of 1 to 6 carbon atoms,alkylamino of 1 to 6 carbon atoms, haloalkyl of 1 to 6 carbon atoms,hydoxyalkyl of 1 to 6 carbon atoms, lower alkoxy of 1 to 6 carbon atomsand alkoxyalkyl of 2 to 12 carbon atoms, R₈ is selected from the groupconsisting of hydrogen, alkyl of 1 to 6 carbon atoms, hydroxyalkyl of 1to 6 carbon atoms, --NH₂, alkylamino of 1 to 6 carbon atoms,alkylaminoalkyl of 2 to 12 carbon atoms, aminoalkyl of 1 to 6 carbonatoms, cycloalkyl of 3 to 7 carbon atoms, cycloalkylalkyl of 4 to 13carbon atoms, alkenyl of 2 to 6 carbon atoms, alkoxy of 1 to 6 carbonatoms, alkoxyalkyl of 2 to 12 carbon atoms, haloalkyl of 1 to 6 carbonatoms and unsubstituted or substituted aryl and arylalkyl of 1 to 6alkyl carbon atoms, the substituents being at least one member of thegroup consisting of alkyl of 1 to 6 carbon atoms, halogen, --NO₂, --NH₂,alkylamino of 1 to 6 carbon atoms, haloalkyl of 1 to 6 carbon atoms,hydroxyalkyl of 1 to 6 carbon atoms, alkoxy of 1 to 6 carbon atoms andalkoxyalkyl of 2 to 12 carbon atoms, R₉ is selected from the groupconsisting of hydrogen, alkyl of 1 to 6 carbon atoms, haloalkyl of 1 to6 carbon atoms, aryl and aryl substituted with at least one member ofthe group consisting of the group consisting of halogen, alkyl of 1 to 6carbon atoms, --NO₂, --NH₂, alkylamino of 1 to 6 carbon atoms, haloalkylof 1 to 6 carbon atoms, hydroxyalkyl of 1 to 6 carbon atoms, alkoxy of 1to 6 carbon atoms and akloxyalkyl of 2 to 12 carbon atoms, R₁₀ isselected from the group consisting of hydrogen, alkyl of 1 to 6 carbonatoms, haloalkyl of 1 to 6 carbon atoms, alkoxy of 1 to 6 carbon atomsand unsubstituted or substituted aryl with the substituents being atleast one member of the group consisting of alkyl of 1 to 6 carbonatoms, haloalkyl of 1 to 6 carbon atoms, hydroxy alkyl of 1 to 6 carbonatoms and a alkoxyalkyl of 2 to 12 carbon atoms, R₁₈ and R₁₉ areindependently selected from the group consisting of hydrogen, halogen,alkyl of 1 to 6 carbon atoms, alkoxy of 1 to 6 carbon atoms and --OH,R₂₀ is hydrogen or halogen, m is an integer from 0 to 6, n is 1 or 2 andtheir non-toxic, pharmaceutically acceptable acid addition salts.
 2. Acompound of claim 1 wherein R₁ is selected from the group consisting ofalkyl of 1 to 6 carbon atoms, alkenyl of 2 to 6 carbon atoms, haloalkylof 1 to 6 carbon atoms and alkoxyalkyl of 2 to 12 carbon atoms, R₂ R₃and R₄ are independently selected from the group consisting of hydrogen,halogen, haloalkyl of 1 to 6 carbon atoms, alkyl of 1 to 6 carbon atoms,--NO₂, --CONH₂, amidoalkyl of 1 to 6 carbon atoms, hydrazino,hydrazinoalkyl of 1 to 6 carbon atoms, azido azidoalkyl of 1 to 6 carbonatoms, ##STR26## substituted or unsubstituted ##STR27## R₅ is selectedfrom the group consisting of hydrogen, halogen, alkyl of 1 to 6 carbonatoms, haloalkyl of 1 to 6 carbon atoms, alkoxy of 1 to 6 carbon atoms,alkoxy alkyl of 2 to 12 carbon atoms, alkylthioalkyl of 2 to 12 carbonatoms, hydroxyalkyl of to 6 carbon atoms, ##STR28## substituted orunsubstituted ##STR29## R₆ and R₇ are individually selected from thegroup consisting of hydrogen, alkyl of 1 to 6 carbon atoms, hydroxyalkylof 1 to 6 carbon atoms, alkylaminoalkyl of 2 to 12 carbon atoms,aminoalkyl of 1 to 6 carbon atoms, cycloalkyl of 3 to 7 carbon atoms,cycloalkylalkyl of 4 to 13 carbon atoms, alkoxyalkyl of 2 to 12 carbonatoms, haloalkyl of 1 to 6 carbon atoms, aryl and aralkyl of 1 to 6alkyl carbon atoms, R₈ is selected from the group consisting ofhydrogen, alkyl of 1 to 6 carbon atoms, hydroxyalkyl of 1 to 6 carbonatoms, alkylamino of 1 to 6 carbon atoms, alkylaminoalkyl of 2 to 12carbon atoms, aminoalkyl of 1 to 6 carbon atoms, cycloalkyl of 3 to 7carbon atoms, cycloalkylalkyl of 4 to 13 carbon atoms, alkenyl of 2 to 6carbon atoms, alkoxy of 1 to 6 carbon atoms, alkoxyalkyl of 2 to 12carbon atoms, haloalkyl of 1 to 6 carbon atoms and aryl and arylalkyl of1 to 6 alkyl carbon atoms, R₉ is selected from the group consisting ofhydrogen, alkyl of 1 to 6 carbon atoms and haloalkyl of 1 to 6 carbonatoms, R₁₀ is selected from the group consisting of hydrogen, alkyl of 1to 6 carbon atoms, haloalkyl of 1 to 6 carbon atoms and alkoxy of 1 to 6carbon atoms and R₁₈, R₁₉ and R₂₀ are hydrogen.
 3. A compound of claim 2where R₁ is ethyl.
 4. A compound of claim 3 wherein R₂ and R₃ areindependently selected from the group consisting of hydrogen, alkyl of 1to 6 carbon atoms, halogen, haloalkyl of 1 to 6 carbon atoms and--(CH₂)_(m) --OR₆ and R₄ and R₅ are independently selected from thegroup consisting of hydrogen, alkyl of 1 to 6 carbon atoms, ##STR30##substituted or unsubstituted --(CH₂)_(n) --N═X wherein said substituentis alkyl of 1 to 6 carbon atoms.
 5. A compound of claim 4 wherein R₄ ishydrogen or ##STR31## R₆ and R₇ are independently hydrogen or alkyl of 1to 6 carbon atoms, R₅ is selected from the group consisting of hydrogen,alkyl of 1 to 6 carbon atoms and optionally substituted--(CH₂)_(n) --N═Xwhen the substituent is alkyl of 1 to 6 carbon atoms and --N═X isselected from the group consisting of piperazinyl, piperidyl andmorpholinyl.
 6. A compound of claim 5 wherein R₂ is hydrogen or halogen,R₃ is selected from the group consisting of hydrogen, halogen, alkyl of1 to 6 carbon atoms and --OR₆ and R₆ is selected from the groupconsisting of hydrogen, alkyl of 1 to 6 carbon atoms and aralkyl of 1 to6 carbon atoms.
 7. A compound of claim 6 wherein R₂ is selected from thegroup consisting of hydrogen, chlorine and fluorine and R₃ is selectedfrom the group consisting of hydrogen, fluorine, chlorine, methyl andmethoxy.
 8. A compound of claim 7 wherein R₄ is hydrogen and R₅ ishydrogen or substituted --(CH₂)_(n) -piperidyl optionally substitutedwith alkyl of 1 to 6 carbon atoms.
 9. A compound of claim 6 wherein R₂is selected from the group consisting of chlorine and fluorine and R₃ isselected from the group consisting of fluorine and methyl.
 10. Acompound of claim 9 characterized in that it is5-ethyl-9,10-difluoro-4,5-dihydro-5-hydroxy-1H-oxepino-[3':4']indolizino[1,2-b]quinoline-3,15[4H,13H]dione.
 11. A compound of claim 4 selected from the group consistingof -5-ethyl-4,5-dihydro-5-hydroxy-1H-oxepino [3',4':6,7]-indolizine[1,2-b] quinoline-3,15 (4H,13H)-dione5,12-diethyl-4,5-dihydro-5-hydroxy-1H-oxepino [3'4':6,7]-indolizine[1,2-b]quinoline-3,15[(4H,13H)-dione-10-benzyloxy-5-ethyl-4,5-dihydro-5-hydroxy-1H-oxepino[3',4':6,7]-indolizino (1,2-b]-quinoline-3,15(4H,13H)-dione-5-ethyl-4,5-dihydro-5,10,-dihyroxy-1H-oxepino[3'4':6,7]-indolizino [1,2-b] quinoline-3,15 (4H,13H)-dione11-(dimethylamino)-methyl-5-ethyl-4,5-dihydro-5-10-dihydroxy-1H-oxepino[3',4':6.7]-indolizino[1,2-b]quinoline-3,15 (4H-13H)-dione5-ethyl-9-fluoro-4,5-dihydro-5-hydroxy-10-methoxy-1H-oxepino[3',4':6.7]indolizino [1,2-b] quinoline-3,15(4H,13H)-dione9-chloro-5-ethyl-4,5-dihydro-5-hydroxy-10-methyl-1H-oxepino[3'4':6.7]indolizino [1,2-b] quinoline-3,15(4H,13H)-dione5-ethyl-9,10-difluoro-4,5-dihydro-5-hydroxy-1H-oxepino [3'4':6,7]indolizino [1,2-b]quinoline-3,15(4H,13H)-dione9-chloro-5-ethyl-4,5-dihydro-5-hydroxy-10-methoxy-4H-oxepino [3'4':6.7]indolizino [1,2-b] quinoline-3,15(4H,13H)-dione5-ethyl-4,5-dihydro-5-hydroxy-10-methoxy-1H-oxepino [3'4':6,7]indolizino [1,2-b]quinoline-3,15(4H,13H)-dione9-11-dichloro-5-ethyl-4,5-dihydro-5-hydroxy-1H-oxepino [3'4':6,7]indolizino [1,2-b]quinoline-3,15(4H,13H)-dione5-ethyl-9-fluoro-4,5-dihydro-5-hydroxy-10-methyl-1H-oxepino [3'4':6,7]indolizino[1,2-b]quinoline-3,15(4H,13H)-dione5-ethyl-10-fluoro-4,5-dihydro-5-hydroxy-1H-oxepino [3'4':6,7]indolizino[1,2-b] quinoline-3,15(4H,13H)-dione10-chloro-5-ethyl-4,5-dihydro-5-hydroxy-1H-oxepino [3'4':6,7] indolizino[1,2-b] quinoline-3,15(4H,13H)-dione10-chloro-5-ethyl-9-fluoro-4,5-dihydro-5-hydroxy-1H-oxepino [3'4':6,7]indolizino [1,2-b] quinoline-3,15(4H-13H)-dione5,12-diethyl-4,5-dihydro-5,10 dihydroxy 11 morpholino methyl-1H-oxepino[3'4':6,7] indolizino [1,2-b] quinoline 3,15(4H,13H) -dione5,12-diethyl-9-fluoro-4,5-dihydro 5 hydroxy-10-methoxy-1H-oxepino[3'4':6,7] indolizino [1,2-b] quinoline-3,15(4H-13H)-dione5-ethyl-4,5-dihydro-5-hydroxy-12-methyl-1H-oxepino [3'4':6,7] indolizino[1,2-b] quinoline-3,15(4H,13H)-dione9-chloro-5-ethyl-4,5-dihydro-5-hydroxy-10-methoxy-12-(4-methylpiperazinomethyl)-1H-oxepino [3'4':6,7] indolizino [1,2-b]quinoline-3,15(4H,13H)-dione9-chloro-5-ethyl-4,5-dihydro-5-hydroxy-10-methoxy-12-morpholinomethyl-1H-oxepino[3'4':6,7] indolizino [1,2b] quinoline 3,15(4H,13H)-dione5-ethyl-4,5-dihydro-5-hydroxy-12-(4-methyl piperazinomethyl)-1H-oxepino[3'4':6,7] indolizino [1,2-b] quinoline-3,15(4H,13H) -dione5-ethyl-4,5-dihydro-5-hydroxy-12-piperidinomethyl-1H-oxepino [3'4':6,7]indolizino [1,2-b] quinoline-3,15 (4H,13H)-dione5-ethyl-4,5-dihydro-5-hydroxy-12-morpholinomethyl-1H-oxepino [3'4':6,7]indolizino [1,2-b] quinoline-3,15(4H,13H)-dione5-ethyl-10-fluoro-4,5-dihydro-5-hydroxy-12-(4-methylpiperazinomethyl)-1H-oxepino[3'4':6,7] indolizino [1,2-b] quinoline-3,15(4H,13H)-dione5-ethyl-10-fluoro-4,5-dihydro-5-hydroxy-12-morpholinomethyl-1H-oxepino[3'4':6,7] indolizino [1,2-b]quinoline-3,15 (4H,13H)-dione5-ethyl-9-fluoro-4,5-dihydro-5-hydroxy-10-methyl-12-(4-methylpiperazinomethyl)-1H-oxepino[3'4':6,7] indolizino [1-2b] quinoline-3,15(4H,13H)-dione5-ethyl-9-fluoro-4,5-dihydro-5-hydroxy-10-methyl-12-morpholinomethyl-1H-oxepino[3'4':6,7] indolizino [1,2-b] quinoline-3,15(4H,13H)-dione5-ethyl-9-fluoro-4,5-dihydro-5-hydroxy-10-methyl-12-piperidinomethyl-1H-oxepino[3'4':6,7] indolizino [1-2b] quinoline3,15(4H, 13H) -dione9-chloro-5-ethyl-10-fluoro-4,5-dihydro-5-hydroxy-12-morpholinomethyl-1H-oxepino[3'4':6,7] indolizino [1-2b] quinoline-3,15(4H,13H)-dione and theirnon-toxic, pharmaceutically acceptable salt.
 12. A method of inhibitingto poisomerase in warm-blooded animals comprising administering towarm-blooded animals an amount of a compound of claim 1 sufficient toinhibit to poisomerase activity.
 13. A method of inhibiting tumor inwarm-blooded animals compound comprising administering a tumor growthinhibiting amount of a of claim 1.